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Fig. I. — Photograph of agar plate prepared with vaccine material 
immediatelv after Glycerination. 

Fig. 2. — Photograph of similar agar plate prepared with vaccine 
material /<wr weeks after Glvcer I NATION. 







M.A., M.D. Cantab., ]$r.R.C.P. Lond. '."Z- 











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Hi story of sm allpox — Inoculation first introduced into England — 
Method of^-Fee for — Vaccination first discovered by Jenner — 
His investigations — Pathology of vaccinia — Cow-pox — Horse- 
pox or "grease" — Cow-pox in the cow — Casual cow-pox in 
man — Inoculated cow-pox in man . . Pages 1-40 



Jenner on vaccinia — Smallpox and cow-pox identical — Variolation 
of bovine animals — Lymph obtained from, used on the human 
subject — Retro- vaccination . . . .41-66 



Jenner's first vaccination — Arm-to-arm vaccination — Origin of lymph 
first used on the Continent and in other foreign countries — 


Lymph of equine origin — Origin of lymph used at the Govern- 
ment Animal Vaccine Establishment — Variola - vaccine 
lymph ...... Pages 67-72 



Cause of papular stage of vaccination — Description of vesicle — 
Action of leucocytes— Pustular stage — Gustav Mann's investiga- 
tions as to changes produced in skin of calf by vaccination — 
Chemistry and morphology of vaccine lymph . . 73-84 



Granules in vaccine lymph — Micro-organisms in — Specific micro- 
organism of — Action of glycerine on — Experiments with horse- 
pox lymph — " Extraneous " micro-organisms in vaccine lymph — 
Tubercle bacillus in — Variolation of bovine animals — Cultivation 
of micro-organisms of variola and vaccinia in artificial media — 
Protozoa . . . . . .85-127 



Investigations as to efficiency of lymph stocks by variolation and 
vaccination of the monkey — Difference between variolation and 
vaccination of the monkey — Comparison of effects produced by 
human and by calf lymph — The " variolous test " . 128-133 




Vaccination with sterilised lymph — Possibility of obtaining tem- 
porary protection by — No micro-organisms found in plate 
cultivations of — Experiments on calves with — Subcutaneous 
inoculation of monkeys and horses with — Immunity conferred 
by serum from previously vaccinated calves . Pages 134-141 



Definition of — History of introduction of — Retro-vaccination — 
Animal vaccination on the Continent and in England — Grounds 
on which it has been advocated — English Royal Commission 
on . . . . . . . 142-152 



Bacteriological purification and preservation of vaccine lymph by 
glycerine — " Extraneous " micro-organisms in vaccine lymph — 
Quantity of vaccine material increased by addition of glycerine — 
Use of glycerinated calf lymph on the Continent — Investigations 
as to action of glycerine on pathogenic and non-pathogenic 
micro-organisms — Special investigations as to its action on the 
tubercle bacillus — English Royal Commission on quality of 
vaccine lymph — Outbreak of disease, Riigen, following use of 
vaccine lymph mixed with thymol and glycerine — Value of 


glycerine, lanoline, and vaseline in the preservation and purifica- 
tion of vaccine lymph compared — Advantages of glycerinated 
calf lymph — Preparation of — Recommendations as to the use 
of, in England .... Pages 153-188 


The preparation and storage of glycerinated calf lymph in certain 
European countries ..... 189-236 


The bacteriology of vaccine lymph . . . 237-242 

INDEX 243-257 



Photographs of agar plate-cultivations of gly- 
cerinated vaccine pulp, before and after 
its storage ..... To face Title-page 


Photograph of case of casual horse-pox in man To face page 32 


Photographs of agar plate-cultivations of vase- 
line vaccine pulp, before and after its 
storage . . . . . ,, ,, 176 


Photographs of agar plate - cultivations of 
lanoline vaccine pulp, before and after its 
storage ..... »» ,, 178 


Photographs of agar plate-cultivations of gly- 
cerinated vaccine pulp, showing, week by 
week, the gradual purification of the lymph 
from extraneous micro-organisms . . >» >> 184 



Photograph of child, vaccinated with glyceri- 
nated lymph free from extraneous micro- 
organisms ..... To face page i86 


Lymph-mixing machine of Dr. Doring, Berlin ,, ,, 214 


Lymph -mixing machine of Dr. Chalybaus, 

Dresden ..... ,, ,, 222 



Just one hundred years ago, in the summer of the 
year 1798, there was pubUshed a treatise, the appear- 
ance of which marked a never-to-be-forgotten event 
in the history of scientific medicine. This truly epoch- 
making pubUcation was entitled "An Inquiry into 
the Causes and Effects of the Variolse Vaccinae, a 
disease discovered in some of the western counties of 
England, particularly Gloucestershire, and known by the 
name of the Cow-pox." It- was the work of Edward 
Jenner, a country medical practitioner, living at Berkeley 
in the vale of Gloucester. 

But in order adequately to appreciate the far-reach- 
ing importance of this work, first issued in the com- 
paratively small compass of a quarto pamphlet of 
seventy-five pages, it is necessary for us, if possible, to 
transport ourselves in thought backwards for over a 
century, in order to reahse approximately the truly 
terrible and far-reaching nature of the scourge, the only 
efficient preventative of which was first introduced to 



the human race through the publication of Jenner's 

It would be difficult to give a more vivid illustration 
of the state of affairs previously existing than is afforded 
by Macaulay's oft-quoted passage, in which reference is 
made to the death in 1694 of Queen Mary, wife of 
William III. He writes-^ 

" That disease, over which science has since achieved 
a succession of glorious and beneficent victories, was 
then the most terrible of all the ministers of death. The 
havoc of the plague had been far more rapid ; but the 
plague had visited our shores only once or twice within 
living memory ; and the smallpox was always present, 
filling the churchyards with corpses, tormenting with 
constant fears all whom it had not yet stricken, leaving 
on those whose lives it spared the hideous traces of its 
power, turning the babe into a changeling at which the 
mother shuddered, and making the eyes and cheeks of 
the betrothed maiden objects of horror to her lover." 

The ever-present danger, and the lively apprehensions 
engendered through recognition of the sHght chance of 
any one evading the onslaught of the scourge between 
childhood and maturity, is also brought out somewhat 
forcibly in an extract from the " Pages from a Private 
Diary," in the January number of the Cornhill Magazine 
for the present year. It runs as follows — " I came on 
a curious passage in a letter of Mrs. Waller's to her 
banished son, about the marriage of his daughter. She 
wishes to know what dowry he is prepared to give. ' I 
am not in haste to marry her, she is young enough to 


stay, but the danger is if she should catch the small- 
pox, or her beauty should change, it would be a great 
loss to her.' Everybody," adds the writer, "is familiar 
with the frequent references to smallpox in the letters 
and memoirs of the seventeenth century. Pepys is full 
of it ; but I have never met with a passage that brings 
so keenly home to me the nearness of the risk." 

In this connection also may be mentioned a record 
of an epidemic of smallpox in the small town of Ware 
in 1722. The population at the commencement of the 
epidemic was 2515, of whom 1601 are stated to have 
had smallpox previously, the remaining 914 being 
therefore presumably susceptible to the disease. During 
the course of the epidemic, of these 914 persons 612 
were attacked, of whom 72 died. The remaining 302 
persons who escaped attack are spoken of in the record 
in quaintly dogmatic fashion as "to have the smallpox." 

We possess also a record of an epidemic which 
occurred at Warrington in 1773, where out of 473 
deaths from all causes during the year, among a popula- 
tion estimated at 8000, no less than 211 were attributed 
to smallpox. And from a report by Haygarth of 
Chester, we learn, as in the case of Ware, how small a 
proportion of the population in these places had not 
suffered from smallpox at one time or another. The 
actual number of persons attacked at Chester in 1774 
was 1202, of whom 202 died, the total population of 
the town being at that time 14,713. But as the result 
of an investigation carried out at the commencement of 
the following year, it becomes fairly certain that, previous 


to the outbreak in 1774, there were in Chester of the 
whole population only 2262, or 15 per cent., who had 
not already had the disease. Consequently, of the re- 
maining susceptible portion of the inhabitants more 
than half were attacked before the end of the year. 
These epidemics at Chester and Warrington further 
illustrate a marked characteristic of smallpox in the 
eighteenth century which is also noticeable in the 
records of preceding periods in which the age of those 
attacked is given, namely, the large proportion of the 
deaths contributed by children under ten years old. 
In the Warrington outbreak of 1773, and also in 
that which appeared in Chester in the following year, 
not a single death occurred in persons above this age, 
while of the 202 deaths at Chester, no less than a quarter 
of the total number were accounted for by infants of less 
than twelve months. Other instances of a like condition 
of affairs, noted in the Final Report of the Royal Com- 
mission on Vaccination, refer to Kilmarnock, where out of 
622 deaths, occurring between 1728 and 1763, of which, 
with nine exceptions, the ages are recorded, only seven 
were of those above ten years, and to records obtained 
from the burial registers of the graveyards of St. Cuth- 
bert's, Canongate, and Buccleuch Street, Edinburgh, from 
which we learn that, during the years 1764-83, of every 
_thousand deaths fr<?m smallpox at all ages, no less than 

.993 were of children below ten years of age. 
/ In considering the question of ratio of deaths from 
L smallpox to the living population we are beset with 
several difficulties. In the first place, as McVail points 


out, in no single epidemic in London in former centuries 
was it possible to see smallpox at its worst, since 
epidemics followed one another at such rapid intervals 
jthat of necessity the material on which at any one period 
their energy could exert itself was extremely Hmited. 
" At any given date the great bulk of the people had 
already had smallpox, and it was only among a fraction 
that there was room for the disease to spread. Yet it 
was among such fractions of the population that small- 
pox every few years slew its thousands in London alone." 
If we desire to obtain an illustration of the full effects 
of an invasion of smallpox, when not thus hampered by 
the existence of very general immunity, the result of 
former epidemics, we must turn our attention to some 
particular locahty which, owing to its isolated position, 
has previously escaped invasion. In illustration of this 
point we may take the case of Iceland, where, during 
1 707-1 709, out of a total population of 50,000, no less 
than 18,000 persons died of this disease. 

Concerning the regions whence smallpox was origin- 
ally derived, and the routes along which it subsequently 
spread, comparatively little accurate information is avail- 
able. That the disease is of great antiquity in China and 
India is generally admitted, but it is fairly certain that it 
was not indigenous in Europe, where it appears to have 
been unknown prior to the end of the sixth century a.d., 
when the disease not improbably extended westwards 
from Arabia, it having broken out in a virulent form 
in the Abyssinian army of Abraha, which was besieging 
Mecca about 569 a.d. Hippocrates and Celsus were 


apparently ignorant of its characteristics, and the same is 
probably true of Galen, although the Arabian physician, 
Rhazes, quotes three passages from Galen's works " Kara 
ykvo%' " On Pulses," and " On the Use of the Members," 
which he considers to prove that the older observer was 
not without knowledge of the disease. 

Rhazes, in his work, quotes from the Pandects of 
Ahron of Alexandria, who flourished in the time of the 
Emperor HeracHus (610-641 A.D.), passages in which 
smallpox is undoubtedly referred to. Another writer, 
almost a contemporary of his own, also mentioned by 
Rhazes as having written a description of the disease, 
is Messua the Elder, who died in 857 a.d. The fact 
that at these dates smallpox was not of recent intro- 
duction is evidenced by the circumstance that no note 
to this effect is made by any of these writers, as other- 
wise might be expected to be the case. 

To Gregory of Tours, who wrote in 581 a.d., we owe 
what is possibly a description of the first outbreak of 
smallpox on the continent of Europe, since in his 
description, " De lue quae cum dessicis fuit," he defin- 
itely differentiates the disease in question from the 
plague with which it was in early days frequently 

It was, in all probability, an extension to France and 
Italy of the epidemic described by Gregory, which has 
been handed down by the writings of Marius, Bishop of 
Averche, who, in the course of his description, makes the 
statement that "this year (570) a violent fever with flux 
of the bowels and variola affected both Italy and France." 


Concerning this statement it is of interest to note that 
we have here what may be regarded as the earliest 
employment of the term " variola " in its modern sense. 
In his Life of Jenner, Baron states, however, that " The 
first recorded case of smallpox or variola under that 
peculiar and now appropriate name is probably that of 
Elfrida, daughter of our English Alfred and wife of 
Baldwin the Bald, Earl of Flanders : date of her illness 
is generally fixed as 907 a.d. ; she recovered." The 
term " variola " has been stated to represent the diminu- 
tive form of the late Latin varus^ a pimple, or, on the 
other hand, it may be derived from varius, meaning 
"spotted." By some authorities it is believed to have 
been first used in its modern sense by Constantinus 
Africanus in the eleventh century. The Anglo-Saxon 
equivalent of variola is found in the word pocca, meaning 
a bag or sack, which has given rise to the modern pock. 
The first mention of the term pocca would appear to be 
in a tenth century Leech-Book of the physician Bald ; 
and subsequently it is employed in the Chronicon Ber- 
tinianurn (1440 a.d.), wherein is set out an account of 
the illness of Baldwin, son of the Earl of Flanders, who 
died of smallpox in 961, the disease being described as 
"variolas sive poccas." Previously to the date of this 
and certain earlier writings, of which mention has been 
made, the word variola had apparently been used some- 
what indefinitely as descriptive of any eruptive disease, 
inclusive of the plague. This was no doubt in part due 
to the defects of diagnosis, the study of medicine being 
at so low an ebb during the Middle Ages. In a reverse 

VA CCINA TION : chap. 

manner the Anglo-Saxon equivalent pocca or pock has, 
since the appearance of syphilis in Europe, become 
restricted in its meaning, so as to designate the eruption 
peculiar to this disease alone. For this reason there 
gradually came about the use of the distinctive term 
S7naIl-^ox, or its French equivalent petite verole, as 
opposed to the large pock, or great pock, now exclusively 
reserved for syphilis. 

To Rhazes himself, who flourished about the year 900 
A.D., and who held the post of physician at the hospital 
at Bagdad, we owe what is really the first scientific 
account of the symptoms of smallpox, although he 
appears to have confounded the disease with measles. 
The treatise of Rhazes was printed in the original Arabic, 
together with a translation into Latin by Channing, in 
1 766, but it was not until 1847 that, under the auspices of 
the Sydenham Society, Greenhill published his English 
translation of this remarkable work. The descriptions 
of the various forms in which smallpox may present 
itself given by Rhazes are clear and definite, and the 
treatment advocated by him is in many respects in 
harmony with that which is now considered most 
appropriate. In support of this statement, it may be 
worth while to quote certain passages from his Treatise 
on the Smallpox and Measles — 

"As to the moderns, although they have certainly 
made some mention of the treatment of the smallpox 
(but without much accuracy and distinctness), yet there 
is not one of them who has mentioned the cause of the 
existence of the disease, and how it comes to pass that 


hardly any one escapes it, or who has disposed the. 
modes of treatment in their right places. 

" Now the smallpox arises when the blood putrefies 
and ferments, so that the superfluous vapours are thrown 
out of it, and it is changed from the blood of infants, 
which is like must, into the blood of young men, which 
is like wine perfectly ripened ; and the smallpox itself 
may be compared to the fermentation and the hissing 
noise which takes place in must at that time. And this 
is the reason why children, especially males, rarely 
escape being seized with this disease. 

"The smallpox arises from a fermentation that 
takes place in the blood, when it is undergoing the 
change from that of an infant to that of a young man ; 
and it is accompanied by great heat and a disagreeable 
odour ; and this is when the patient is of a hot tempera- 
ment. The disease sometimes happens twice or even 
three times to the same individual, but generally once, 
especially to males, for the blood of infants must 
necessarily undergo this change. 

" I am now to mention the seasons of the year in 
which the smallpox is most prevalent : which are, the 
latter end of the autumn and the beginning of the 
spring ; and when in the summer there are great and 
frequent rains with continued south winds, and when 
the winter is warm and the winds southerly. 


"The eruption of the smallpox is preceded by a 
continued fever, pain in the back, itching in the nose, 
and terrors in sleep. These are the more peculiar 
symptoms of its approach, especially a pain in the back, 
with fever ; then also a pricking, which the patient feels 
all over his body ; a fulness of the face, which at times 
goes and comes \ an inflamed colour, and vehement 
redness in both the cheeks ; a redness of both the 
eyes ; a heaviness of the whole body ; great uneasiness, 
the symptoms of which are stretching and yawning ; a 
pain in the throat and chest, with a slight difficulty in 
breathing, and cough ; a dryness of the mouth, thick 
spittle, and hoarseness of the voice ; pain and heaviness 
of the head. 

"When, therefore, you see these symptoms, or some 
of the worst of them (such as the pain of the back, and 
the terrors in sleep, with the continued fever), then you 
may be assured that the eruption ... in the patient 
is nigh at hand. 

" Ibn Masawaih says — ' When there is with a con- 
tinued fever, a redness of the eyes and face, much 
heaviness of the body and head, itching of the nostrils, 
sneezing, and a pricking pain all over the body, these 
are signs of the eruption of the smallpox. Your first 
care should be directed to the eye, for which you should 
use a collyrium made of sumach and rose-water, in 
order to prevent any pustules from coming out in it. 


"The eruption of the smallpox ... is accelerated 
by well wrapping the patient up in clothes, and rubbing 
his body, by keeping him in a room not very cold, and 
by sipping cold water, a Httle at a time, especially when 
the burning heat is very great ; for cold water, when it 
is sipped a little at a time, provokes sweat, and assists 
the protrusion of the superfluous humours to the surface 
of the body. 

"Extinguishing remedies are to be used when you 
see that, as soon as any of the pustules come out and 
appear in the skin, the patient finds himself easier after 
it, and his pulse and breathing are relieved in propor- 
tion ; but if you see that the eruption and appearance 
of the pustules goes on slowly and with difficulty, you 
must in this case avoid all very extinguishing medicines, 
for to use them would be acting contrary to Nature, 
and hindering her from throwing out the superfluous 
humours upon the skin. 

" As soon as the symptoms of the smallpox appear, 
we must take especial care of the eyes, then of the 
throat, and afterwards of the nose, ears, and joints, in 
the way I am about to describe. And besides these 
parts, sometimes it will be necessary for us to extend 
our care to the soles of the feet, and the palms of the 
hands ; for occasionally violent pains arise in these 
parts from the eruption of the smallpox in them being 
difficult on account of the hardness of the skin. 


" If a severe pain arises in the soles of the feet, then 
take care to anoint them with tepid oil, and foment 
them with hot water and cotton : ... for these and 
the like things soften and relax the skin and thus 
faciUtate the eruption of the pustules, and lessen the pain. 

"All those pustules that are very large should be 
pricked ; and the fluid that drops from them be soaked 
up with a soft clean rag in which there is nothing that 
may hurt or excoriate the patient. 

" When the desiccation of the pustules is effected, 
and scabs and dry eschars still remain upon the body, 
examine them well, and upon those that are thin and 
perfectly dried up, and under which there is no mois- 
ture, drop warm oil of sesamum every now and then, 
until they are softened and fall off. 

"... and in order to efface the pock-holes, 
and render them even with the surface of the body, let 
the patient endeavour to grow fat and fleshy, and use 
the bath frequently, and have his body well rubbed." 

It appears certain that during the time of the Crusades 
smallpox spread in a manner that has not unfrequently 
been witnessed subsequently under similar circumstances 
of war prevalence ; the numbers of those dying from the 
scourge during this period, as far as can be learned from 
the indefinite accounts now available, having apparently 
been extraordinarily great. There is reason also for the 


supposition that it was during the epidemic following 
the Crusades that smallpox houses were first erected. 
There is some authority for thinking that smallpox first 
found its way into England in the year 1241-42, although 
it may be stated that Creighton considers that there is no 
trustworthy evidence of its existence in England earlier 
than the sixteenth century. Towards the close of the 
thirteenth century the disease was referred to by Gil- 
bertus Anglican us in his Compendium Medicince, which is 
probably the earliest medical work produced in England. 
But although he gives an account of the symptoms of 
smallpox and measles, he was not, so far as can be 
judged, himself acquainted with the disease, and he did 
no more than slavishly follow the early accounts of the 
Arabian physicians. The same statement applies also 
to his successor, John of Gaddesden, physician to 
Edward II., whose work, Rosa Anglica^ appeared 
about the year 1320. He gives a description of an 
attack of smallpox from which the King's son ^ suffered. 
For this, Gaddesden treated him by wrapping him in a 
red cloth, and by entirely enclosing the bed in hangings 
of a red colour, with the effect, he says, that no pitting 
of the skin resulted from the attack. This method of 
treatment, however, did not originate with Gaddesden, 
having been suggested by former writers of the Arabian 
schools, to whom reference has already been made. 
Baron says of Gaddesden that " his only dogma worth 
recording with regard to smallpox is 'ahquando variolae 

^ Dr. Norman Moore supposes the "King's son" to have been 
Thomas of Brotherton. 



bis hominem invadunt,' " and Sir Thomas Watson some- 
what unreasonably opines him to have been a sorry 

The use of red hangings and wraps was devised by 
its earliest advocates with the object of drawing the 
peccant humours of the body to the surface, while 
puncture of the pustules by means of a golden needle, 
or even a thorn, was relied on for the prevention of 
pitting. That Gaddesden knew of this latter method is 
obvious from his writings, but he does not make any 
mention of having practised it on the King's son. For 
this reason Creighton expresses the opinion that the 
disease, concerning which the old physician says that he 
obtained a good cure, and sine vestigiis, was in all prob- 
abihty something of a much less formidable nature than 

Creighton further asserts that it is not until the year 
1 5 14 that a definite reference to smallpox is to be 
found, this being in a letter in which mention is made 
of an illness from which the king, Henry VIII., had 
recently recovered, ^^ nommee la petitte verolle.''^ The 
English equivalent of this term, however, is shown by 
the researches of Dr. Norman Moore to have been in 
use at a considerably earlier date. The work cited by 
Dr. Norman Moore is a manuscript copy of the 
Breviarium BartholomcBi^ compiled by John Mirfield, 
a canon regular of St. Austin in the priory of St. 
Bartholomew in West Smithfield, and written about 
1387 for the hospital of St. John the Baptist, attached 
to the Abbey of Abingdon. On folio 43^ of the 


manuscript, which is now in the library of Pembroke 
College, Oxford, after the heading " De variolis et 
morbilis " the words, "/.^. smal pockes " are written. 

During the sixteenth century, references to attacks 
by the disease occur with gradually increasing frequency. 
It would appear, however, from the perusal of such 
records as are available, that the disease was not for 
the most part of extremely virulent type, and also that 
its power of infectivity had not as yet been sufficiently 
appreciated to cause it to be feared, at any rate, as was 
the plague, for instance. In support of this assertion 
may be mentioned a death-bed statement by a certain 
Master Richard Allington, recorded by John Stow as 
having been made in the presence of the Master of the 
Rolls and four other eminent lawyers, in which the 
following passage occurs : — 

" Maisters, seinge that I muste nedes die, which I 
assure you I nevar thought wolde have cum to passe 
by this dessease, consyderinge it is but the small 
pockes. ..." 

In 1604, the classification of deaths in London by 
the Company of Parish Clerks was commenced, although 
their annual and weekly records were not regularly 
printed until a quarter of a century later. During this 
interval smallpox made rapid progress in England, 
the Hterature of this and succeeding periods now teem- 
ing with allusions to the "cruel and impartial sickness." 
Dr. Gee, in an address to the Abernethian Society, 
G '.otes numerous instances : — " Even the poets could 
not avoid the disgusting theme. If they wished to 


bewail the death of a friend, in all probability he died 
of the smallpox. Dryden wrote elegies upon two of 
its victims. The first was Lord Hastings, who died, 
1649, ^t ^^ ^g^ °f nineteen. Mrs. Anne Kelligrew 
died of variola in 1685 at the age of twenty-five, and 
she attained the honour of being celebrated by the 
same poet in much nobler verses. Mrs. Katherine 
Phillips, 'the Matchless Orinda,' died of smallpox in 
1664 at the age of thirty-three; she was lamented by 
Cowley. In 1675 Oldham devoted an ode of extra- 
ordinary length to the memory of his friend, Mr. 
Charles Morwent, who was carried off by smallpox. 
Writers of a satirical turn condoled with ladies upon 
the sad loss of beauty which ensued when they escaped 
with life from the dreadful pest. Verses were written, 
' Upon a Gentlewoman, whose nose was pitted with 
the Smallpox,' and so on." 

It was during the epidemic of 1694 that Queen 
Mary, the wife of William III., who was then only 
thirty-three years old, became attacked with what 
eventually turned out to be haemorrhagic smallpox. 
To this attack she succumbed on the eighth (ninth ?) 
day of the disease. The number of deaths from small- 
pox recorded in London alone during this year was 
not far short of 1700, but this figure had been exceeded 
at least seven times during the previous half century, 
most markedly so in 1681, in which year no less 
than 2982 deaths occurred in London from this 

From 1695 onwards for a series of years, the mor- 


tality from smallpox underwent considerable diminu- 
tion j but in 1 7 1 o a terrible epidemic accounted for 
no less than 3138 deaths in London, and we learn, 
on the authority of Blomefield, that the disease also 
cut off "great numbers in Norwich." The year 17 14 
was also marked by epidemic prevalence of smallpox 
in London, which resulted in an almost equal number 
of deaths; while the record of 17 10 was even exceeded 
in 1 7 19. Beyond this period it is needless to extend 
our review, seeing that the history of smallpox pre- 
valence now becomes intimately bound up with the 
history of the spread of certain prophylactic measures, 
the effect of each of which on the disorder will require 
separate consideration. ^^ ^-^^ P-^ t/rcM/^ 1 ^? 

Aitken calls attention to the fact thajTsince the date 


of the first accounts by the Arabi arr-physicians of the 
ravages of smallpox m Mecca , the history of this disease 
may be arranged in three great stages, each of which is 
characterised by remarkable epochs. The first of these 
periods is marked by an improvement in the treatment of 
smallpox, the merit of this revolution in medical practice ^ 
bemg—dne-^o Sydenhamr— In few diseases, indeed, has 
medical opinion undergone in the course of years a 
more obviously beneficial change. The second stage is 
marked by the discovery of the singular phenomenon 
that the virulence of the poison of smallpox is greatly 
mitigated by introducing or engrafting the disease into 
the system through the cutaneous tissues, thereby 
causing the transference of the disease from one 
person to another by inoculation. 

c - 


To Lady Mary Wortley Montagu in 1 7 2 1 we owe 
the introduction of the practice of inoculation into this 
country, a deed which must be considered as one of 
no httle bravery, when measured by the knowledge 
possessed by physicians in those days. The third great 
era is characterised by the remarkable discovery, w^hich 
has rendered the name of Jenner immortal, namely, / 
the modifying and protecting influence of vacciph 
ation. — ^^,^__ _ _. 

With the therapeutic treatment of smallpox we are 
not on the present occasion concerned. On the other 
hand, the practice of inoculation, although prohibited 
by law since the passing of the first Vaccination Act, 
1840, is yet deserving of some attention. Having 
dealt with this matter, we shall be in a position more 
fitly to discuss the question of the discovery of vaccina- 
tion, and of the evidence on which its employment was 
originally based. 

There will remain for our consideration a review of 
such accretions to our knowledge of the subject of 
vaccination, more particularly from the standpoint of 
pathology, as the progress of scientific research during 
the century since Jenner wrote has rendered avail- 

To marshal these in order, to try to indicate their 
relative importance, and to direct attention to their 
bearing on the necessity for certain changes in the 
regulation and practice of vaccination, as these obtain 
at the present time, is the chief aim which I have had 
in view in the preparation of these lectures. 



Recognition in former times of the certainty that 
almost every one would suffer attack by smallpox during 
some period of his or her life, and the lively horror 
inspired by the disease, by reason not only of the high 
fatality inseparable from it, but also of the maiming and 
disfiguring effects on a large proportion of those who sur- 
vived its invasion, naturally turned men's minds to the 
question as to whether it were not possible to devise 
some method for mitigating the virulence of this disorder. 

Whence, however, was originally derived, or by 
whom, accidentally or otherwise, was discovered the 
artificial method of communicating smallpox, which 
came to be known by the term "inoculation," it is at 
the present day impossible to determine. 

According to tradition, the process of inoculation has 
been in use among the Brahmins in India for centuries, 
and a similar claim has been made on behalf of the 

There can be little doubt that in China a method of 
artificially communicating smallpox was in vogue long 
ages apparently before the disease had obtained recog- 
nition in Europe; but the process was essentially different 
from inoculation properly so called, consisting as it did of 
inserting smallpox crusts into the nostril of the patient, 
whereby the disease was communicated by way of the 
respiratory tract. Coming to comparatively recent times, 
however, we have definite evidence that inoculation was 
practised in Turkey early in the eighteenth century. 


In the twenty-ninth volume of the Philosophical Trans- 
actions of the Royal Society may be found references to the 
process, as observed in 17 13 by Dr. Emanuel Timoni 
of Athens, and in 1716 by M. Pylarini, then Venetian 
Consul at Smyrna. But it would seem that their 
descriptions made little, if any, impression at the time, 
and, although published in the records of the Royal 
Society, passed almost at once into oblivion. 

The actual introduction of inoculation into England 
was, as just stated, brought about through the instru- 
mentality of Lady Mary Wortley Montagu, who, apart 
from her social status as wife of the English Ambassador 
at the Ottoman Court, has achieved for herself lasting 
recognition in literature, by reason of her attainments 
in the gentle art of letter-writing. 

Indeed, Professor Rayleigh, in a recent lecture at 
the Royal Institution, gave it as his opinion that in this 
respect she should be placed second only to Madame 
Sevigne. Writing from Adrianople, in 171 8, she says : 
" The smallpox, so fatal and so general amongst us, is 
here entirely harmless by the invention of engrafting, 
which is the term they give it. Every year thousands 
undergo the operation, and the French Ambassador 
says, pleasantly, that they take the smallpox here by 
way of diversion, as they take the waters in other 
countries. There is no example of any one who has 
died in it, and you may believe I am well satisfied of 
the safety of this experiment, since I intend to try it 
on my dear little son. I am patriot enough to take 
pains to bring this useful inv^tion into fashion in 


England." Lady Montagu had indeed the courage 
of her opinion, and the first person inoculated in 
England was her daughter. This first operation, by 
Dr. Maitland, in April 1721, was shortly followed by 
the inoculation of a child of her family physician. Dr. 
Keith. Next an experiment on a larger scale was 
carried out on certain condemned felons, who were 
offered the alternative of submitting to the operation. 
No ill result having been witnessed thus far. King 
George L, who had been much interested in the 
matter, was himself inoculated, as also were several 
members of his family. Apparently, however, the 
method did not generally commend itself to public 
taste ; moreover, it was discovered that, contrary to 
anticipation, persons inoculated with smallpox became 
themselves centres of infection ; and perhaps for the 
reason that no serious attempt was made at that time 
to diffuse information on the subject, it was net until 
thirty years, or more, subsequent to its introduction 
into this country, that inoculation as a prophylactic 
against smallpox became practised to any considerable 

In his Synopsis Medicince, published in 1761, Dr. 
John Allen, a Fellow of the Royal Society, gives the 
following curious directions for the inoculation of the 
smallpox — "First, a little quantity of purulent Matter 
must be procured, about 5 or 6 drops taken from the 
Pustles of some young Person, who has the benign 
Small-Pox of the distinct Kind, which must be brought 
to the place where the operation is to be performed in a 


Small Vial or in a little Pill-Box ; and when all Things 
are prepared, the Surgeon is to make two small Wounds, 
one on the Arm, and the other on the Leg, of the 
Opposite Side ; and the aforesaid variolous Matter is to 
be applied to the Wounds on Dossils of Lint, and a 
Bandage to be made over it. There is no occasion to 
make the incisions deeper than just through the Skin, 
nor any longer than a Barley-Corn. After twenty-four 
Hours the Dossils may be removed, and the Ulcers 
dressed with Diachylum Plaister or Cole-Wart leaves 
once or twice a day, according as more or less Matter 
is discharged. This being done, the Distemper will 
gradually be produced ; and it is usual for the following 
Symptoms to arise. About the eighth day after the 
Operation some Bustles begin to appear not unlike to 
those that are commonly seen in the distinct kinds, a 
little Fever having preceded the Eruption, and the other 
usual Symptoms but more mild and gentle. As to the 
places where the Incisions are made the fourth Day 
they grow red and are inflamed, on the sixth they tend 
to Suppuration, and discharge an ichorous Pus. Indeed, 
about the time of the Eruption of the Bustles, whilst the 
little Fever lasts, the Ulcers discharge somewhat less ; 
but after the Eruption even until the Decline of the 
Pustles, the Pus increases daily, and after that it again 
gradually diminishes ; so that commonly in five weeks' 
time the Ulcers are quite dried up. By how much 
the greater the discharge of purulent Matter is by these 
artificial Ulcers so much th.* milder will the Distemper 
be. In the general it is observable that the Small-Pox 


procured by Inoculation are of the distinct Kind, for the 
most part void of Danger, that the Pustles are few in 
number, and pit very httle. Very often however there 
follow after them Tubercles and Abscesses, some indeed 
but slight ones but sometimes very malignant ones, and 
exceedingly difficult of Cure. 

" To prepare the Body for Inoculation a great many 
talk much of Bleeding, of giving Emeticks, and Canthar- 
ticks. But in truth, if the Person to be Inoculated has 
Youth on his side, is of a good Habit of Body, and in 
Perfect Health, none of these things seem to be neces- 
sary. It is requisite only, that he take care to observe a 
temperate Regimen for some time before the Operation. 
When the Operation is performed, the Person need not 
be kept under any strict Confinement, but may live 
pretty much after his accustomed Manner ; excepting 
that it will be prudent to forbid him the eating any 
Flesh-Meat. When the Small-Pox are come out, the 
same Regimen is to be observed, as is usual in the 
distinct Kind, got in the common way ; and if there 
be occasion for anything at all of Medicine, these 
procured by Art require the same Method of Treat- 
ment, as the benign Discrete Small-Pox in the natural 

The subsequent extensive diffusion of the practice 
of inoculation in this country was undoubtedly due in 
large measure to the labours of two brothers named 
Sutton, and also of Adams and of Dimsdale, of whom 
the last named had a barony conferred upon him in 
commemoration of his having successfully inoculated 

24 VACCINATION: chap. 

the Empress Catherine of Russia. For his services he 
also received a fee of ;^i 0,000, which has been estimated 
as equal to at least three times that sum at the present 
day, and ;^2ooo as an allowance for traveUing expenses. 
He was also appointed Councillor of State and 
Physician to Her Imperial Majesty, with an annuity of 
;^5oo, which was punctually paid him till his death in 
the early years of the present century. 

The Suttons — two brothers, Robert and Daniel, who 
lived at Bury St. Edmunds, Suffolk, and Ingatestone in 
Essex, respectively — although not medical men, intro- 
duced or rather revived the greatly improved method of 
treating the patients under their charge. They were 
extraordinarily successful in the results which they 
obtained, and indeed claim to have inoculated no less 
than 20,000 persons without losing one as the direct 
result of the operation. Acting on the instructions 
originally laid down by Sydenham for the treatment of 
ordinary smallpox — the form of treatment to which, 
by the way, his contemporaries obstinately refused their 
adherence — the Suttons adopted and caused their 
patients strictly to observe what was known as "the 
cooHng treatment," a method much akin to that which 
appears to have been pursued with good results by the 
Turkish operators from whom Lady Montagu first derived 
the knowledge of the process of "engrafting." In the 
hands of Daniel Sutton, who became specially famous 
for his successful inoculations, " the great secret of his 
success seems to have consisted in his making one 
puncture only; exposing his patients much and often 


to a cool atmosphere ; supplying them freely with 
refrigerant drinks; and restricting them to a spare 

By attention to the mode of life and general treatment 
of persons undergoing the process, together with careful 
selection of the sources (preferably the primary vesicle), 
from which the virus was obtained, Adams indeed suc- 
ceeded in gradually evolving a strain of virus of such 
tenuity as to produce in a considerable majority of in- 
stances nothing beyond a single vesicle at the original 
site of puncture. The general eruption, typical of the 
disease as contracted in the ordinary manner, was in these 
cases either entirely absent or represented by, at most, a 
very few secondary vesicles only. Although, doubtless, 
the extremely mild form of the disease, thus artificially 
produced, was none the less contagious, the visible 
effect produced so closely resembled the results then 
beginning to be known as following on the Jennerian 
process of vaccination, that numbers of Adams' patients 
could hardly be persuaded that he had not, contrary 
to their desire, intentionally vaccinated rather than 
variolated them. 

There has always been considerable controversy as 
to the efficacy of the process of inoculation in restrain- 
ing the ravages of smallpox, the matter being rendered 
the more difficult since it is almost impossible to arrive 
with any accuracy at what may be thought of as the 
average death-rate of the natural disease, which in pre- 
inoculation times is known to have varied greatly in 
different epidemics. If, as has been frequently asserted, 

26 VACCINATION : chap. 

the average mortality in natural smallpox may be taken 
as being one in five or six, and, on the supposition 
that in the majority of cases, at any rate, inoculation 
prevented subsequent invasion by smallpox, then it is 
quite evident that the process was of the utmost value 
to the persons inoculated. 

The statement of the Suttons, already mentioned, 
that inoculation in their hands was unattended with 
fatal results, is perhaps exaggerated, as may also be the 
declaration of Baron Dimsdale, that in a series of 
15,000 cases no death had occurred. Neverthe- 
less, the records of the Smallpox Hospital for a series 
of years show that we shall not greatly err, and shall 
indeed be probably within the mark, if we accept 
Dr. Gregory's estimate of i death in 500 inocu- 
lations as approximately correct. There is, however, 
perhaps ground for suspicion that inoculations performed 
in the face of epidemic smallpox were more prone to 
be fatal than were inoculations undertaken when the 
disease was not prevalent. On the other hand, we 
must bear in mind that the general diffusion of inocu- 
lation brought under the influence of the disease, 
although possibly in a mild form, large numbers of 
persons who otherwise might probably have escaped 

It is, I think, hardly possible better to sum up the 
matter than in the words of Sir Thomas Watson : " The 
advantages of the practice of inoculation to the indi- 
vidual, supposing him doomed to have smallpox, were 
great and obvious \ to the community at large they 


were very doubtful. It gave the undoomed individual, 
for certain, an ugly disease, which was comparatively 
free from danger, in exchange for the chances, on the 
one hand, of contracting a very hazardous form, and, on 
the other, of escaping altogether from any form of 
variola. We need not inquire which is the most 
eligible branch of this alternative ; we know which was 
by most men actually chosen. But the practice of 
inoculation, by carrying the virus and the disease into 
every village throughout the length and breadth of the 
land, filled the country with contagion ; ensured the 
disease to all who were subjected to the operation, and 
diminished to all who were not the chances of escaping 
it. No doubt the distemper was produced artificially 
in many more persons than would have caught it 
naturally, had inoculation never been thought of. So 
that while the relative mortality, the percentage of 
death from smallpox, was lessened by this practice the 
absolute mortality was fearfully increased. Such at 
least is the judgment expressed by most who had 
thought and written on the subject." 

As there appear to be few, if any, records as to the 
fees obtained by inoculators, it may be of interest here 
to insert a copy of a small handbill, which is in the 
possession of Mr. Parson, the senior medical prac- 
titioner in Godalming, Surrey. This relates to the 
practice of inoculation, as carried out by a direct 
ancestor and predecessor, in the same practice, of Mr. 
Parson — 

28 VACCINATION : chap. 



R. PARSON, Surgeon, at Has le me re, hav- 
ing provided a convenient Houfe (within a Mile of 
that Town) for the Reception of Patients, propofes 
to inoculate from this Time to Mav next, at ten 
fhillings and fixpence each Perfon. 

lotb^ December, 1783. 

P£T£RSFIELD, Printed by T. WitLMER, 


It was during Jenner's apprenticeship to Mr. Ludlow, 
a medical man practising at Sodbury, near Bristol, that 
his attention first became directed to a belief widely 
prevalent in Gloucestershire during the latter half of 
the eighteenth century, that those persons who in the 
course of their employment on dairy farms happened 
to contract cow-pox, were thereby protected from a 
subsequent attack of smallpox. 

Although himself a native of Gloucestershire, he 
does not appear to have been aware of the local 
tradition, previously to joining Mr. Ludlow at Sodbury. 
It was while working here that his interest became 
aroused by a casual remark made by a young country- 
woman, who happened to come one day for advice. 


and who on hearing mention made of smallpox, 
immediately volunteered the statement that she could 
not take that disease as she had had cow-pox. 

On coming up to London in 1770, to finish his 
medical education, Jenner entered at St. George's 
Hospital, and became also a pupil of John Hunter, 
with whom he speedily became a great favourite. 
With Hunter he discussed the question of the pro- 
tective power of cow-pox over smallpox, but the great 
anatomist does not appear to have attached much 
importance to the matter, although subsequently on 
several occasions he referred to it both publicly and in 
private. In 1773 Jenner returned to his native village 
of Berkeley, where he commenced to practise as a 
medical man. The impression which had been made 
on him in his student days as to the possibiHty of 
obtaining protection against smallpox still persisted, 
becoming even stronger as time went on. It would 
appear, indeed, to have been ever present in his mind, 
and at length he determined to put the matter to the 
test of direct experiment. 

It was not, however, until 14th May 1796, which 
has been appropriately termed the birthday of vaccina- 
tion, that he commenced actual experiment. On this 
day he vaccinated James Phipps, a healthy boy about 
eight years old, by inserting into two superficial in- 
cisions on his arms, matter taken from a vesicle on the 
hand of Sarah Nelmes, a dairymaid, who had been 
infected while milking her master's cows. The result 
of this experiment, the details of which are so well 

30 VACCINATION : chap. 

known as to need no mention here, must have afforded 
Jenner the most lively satisfaction, conforming as it 
did to what, up to that time, had been with him a pious 
opinion only. " But the most agitating part of the 
trial still remained to be performed. It was needful to 
ascertain whether he (Phipps), was secure from the 
contagion of smallpox. This point, so full of anxiety 
to Dr. Jenner, was fairly put to issue on the first of the 
following July." Variolous matter, taken directly from 
a pustule, was carefully inserted by several incisions, but 
no result followed. 

Having succeeded in communicating cow-pox from 
one human being to another by implantation of the 
virus in the skin, the next step was obviously to 
attempt its transference to yet another subject, and so 
on if possible through an indefinite series, in order 
to ascertain whether its protective power was hereby 
diminished. Through lack, apparently, of material, 
Jenner's work was interrupted until the spring of 1798, 
when a simultaneous outbreak of cow-pox and " grease " 
afforded him the further opportunity he desired. 

A few months later he published his Inquiry^ in 
which were set out the results of his observations. 
The appearance of this work was the signal for an 
extraordinary wave of enthusiasm, which spread so 
rapidly that, within a few years only of Jenner's 
announcement, a knowledge of his method of prevent- 
ing smallpox had extended to almost every portion of 
the civilised world. 

The experience of the past century, culminating in 


the definite pronouncement in_ 1896^ of Jthe R oyal 
Commission on Vaccination, having fully justified the^_ 
principle of Jenner as to the prophylact ic power of. 
vaccination, jnna5rbe~of interest to determine what 
light pathology is capable of throwing on the relation- 
ship which may be thought of as existing between 
variola and vaccinia. 

Pathology of Vaccinia 

Vaccinia is, in the human subject, a specific disorder 
characterised by the appearance of a local eruption pass- 
ing through the stages of papule, vesicle, and pustule, 
associated with more or less constitutional disturbance. 

These symptoms are produced indifferently, by the 
inoculation of lymph derived from vesicles similarly 
brought about in a previous case in the human being, 
or from the eruptive vesicles of a disease of bovine 
animals, called cow-pox. Such inoculation process, 
whichever way induced, is known as vaccination. This 
name was originally devised by Mr. Dunnmg of Ply- 
mouth Dock, inspired doubtless by the terminology of 
Jenner, who wrote of the disorder under the title of 
Variolae Vaccinae. / In this manner Jenner gave expres- 
sion to his belief tKat the malady commonly known as 
cow-pox was, in reality, nothing more nor less than 
smallpox of the cow./ 

But soon it was discovered that if there was such a 
malady as "smallpox of the cow," there was also a 
smallpox of the horse, which, under the name of 
"grease," was resorted to as a source of vaccine lymph. 


Jenner, indeed, was undoubtedly of the opinion that 
cow-pox originated from the transmission, most prob- 
ably by human agency, of infection from a previous 
case of horse-pox. At the time of publication of his 
Inquiry^ he had not indeed been able to adduce any 
direct experimental evidence in support of his conten- 
tion, although he had collected a number of observa- 
tions which, in his opinion, afforded reasonable prob- 
ability as to the correctness of his view. 

Mr. Tanner, however, shortly after put the matter 
to the proof, and was successful in communicating the 
disease to the cow, by employing for its vaccination 
lymph taken from the heel of a horse suffering from 
"grease." The attempt resulted in the appearance of 
a perfect vaccine vesicle on the cow's teat, at the site 
of insertion of the lymph. Tanner states that : 
"From handling the cow's teats, I became infected 
myself and had two pustules on my hand, which 
brought on inflammation, and made me unwell for 
several days. The matter from the cow, and from my 
own hand, proved efficacious in infecting both human 
subjects and cattle." 

Dr. Loy was the first to distinguish, in any satisfactory 
fashion, constitutional " grease " from a merely local 
affection, with which it was apt to be confounded ; and 
thus he explained the failure on the part of many 
experimenters to transmit horse-pox to the cow. 

Dr. Sacco of Milan, who, in the first instance, had 
expressed opinions adverse to Jenner's views, subse- 
quently, in a letter written to him in 1803, withdrew 


Casual Horse-Pox. 

George T , stableman, (st. 30, vaccinated in infancy. 

From a photograph taken 6th April 1898 (eighth day of disease) 


them ; his reasons for which may perhaps be best 
given in his own words — 

" Milan, le 25 mars, 1803. 

" Monsieur — J'etais depuis longtemps. occupe a faire 
des experiences sur le grease pour confirmer voire opinion 
sur I'origine de la Vaccine. Jusqu'au commencement 
de cette annee je n'avais jamais pu rien obtenir. La 
lecture du petit livre de Mr. Loy m'encouragea a 
repeter une autre suite. L'hiver de cette annee ne 
pouvait pas etre plus abondante de grease k cause de la 
quantite de I'eau qu'il y avait, et par consequence de la 
boue dans les chemins : ainsi presque tous les chevaux 
souffraient le grease. Mon domestique en fut attaque 
au deux avant-bras par cinq boutons pansant un de mes 
chevaux qui avait le grease ; il ne m'en averti que 
quand les boutons passaient en exsiccation ; celui 
m'encouragea de plus a continuer mes tentatives. J'ai 
inocule plusieurs enfans, plusieurs vaches avec le virus 
qui sortait du grease a differentes epoques, mais toujours 
inutilement. Un cocher se presenta a I'hopital pour 
se faire visiter d'une eruption qu'il avait sur les mains. 
On connait de suite que c'etait vaccine prise en 
traitant les chevaux qu'efifectivement il pansait. II fut 
conduit a I'hopital des enfans trouves oii on fit quelques 
inoculations : il vint le meme jour chez moi, et je fis 
neuf inoculations sur autant d'enfans, et de plus j'ai 
inocule les pis d'une vache. Trois de ces enfans ont 
contract^ une eruption toute pareille k la Vaccine. La 



vache n'a point pris. J'ai fait des autres inoculations 
avec la matibre prise de ces enfans, et c'est dejk la qua- 
tribme generation que se reproduit avec la meme effet 
comme le Vaccin. J'ai dejk inocule plusieurs de ces 
individus avec la petite verole, mais sans aucun effet. 
C'est done bien sur et consent^ que le grease est cause de 
la Vaccine^ et on pouvait bientot changer denomination 
en equine^ ou en ce que vous croyez mieux. J'ai aussi 
enfin obtenu avec le virus de grease inocule sur six 
autres enfans deux boutons tous semblables aux Vaccins. 
Je continue mes observations. II y a tout pour s'assurer 
qu'enfin nous aurons du grease le virus pour se mettre 
\ I'abri de la petite verole sans passer aussi par rinter- 
medium de la vache. J'espbre que cette nouvelle preuve 
pourra oter les doutes qu'il y avaient encore sur I'origine 
de la Vaccine. Je publierai les r^sultats de ces ex- 
periences sur un code doctrinal de vaccination, auquel 
j'ajouterai une planche illumine de grease. J'espbre que 
vous aurez regu les m^dailles par Mr. Woodville, k qui 
je me pris la liberty d'adresser le paquet pour vous le 
faire obtenir avec certitude. Je renouvellais mes remerci- 
mens pour les livres que vous m'avez envoy^s, avec les 
regrets aussi de n'avoir pas re9u aucune de vos lettres. 
Je comte \ cette heure plus de 25 mille inoculations 
faites par moi seul. Je vous prie, mon tr^s-estimable 
collegue, de me donner quelque nouvelle avec quel- 
qu'autre enseignement sur cette matifere, mais surtout 
tout honorez-moi de votre r^ponse. 

Trbs humble serviteur, 

(Sd.) Louis Sacco, Med, Chirurgy 


That a constitutional disease of the horse, character- 
ised by a vesicular eruption, can be induced by the 
inoculation of this animal with the virus of cow-pox or 
vaccinia, has been shown experimentally by Chauveau, 
who injected vaccine lymph subcutaneously and also 
into the blood-vessels and lymphatics, of colts. In 
nearly half the number of cases operated on, the injec- 
tion of the lymph was followed by a generalised eruption 
which Chauveau called "horse-pox." / In all probability 
Jenner was mistaken in his assumption that "grease," 
in the sense of horse-pox, was a necessary antecedent 
to cow-pox ; but at the same time there can be little 
doubt that the two diseases are very closely allied, if 
indeed they be not identical./ 

That this is so is shown by the fact that numerous 
strains of vaccine lymph have, from time to time, been 
raised from the equine source — the protective power of 
which against smallpox we have reason to believe was 
equal to that exhibited by lymphs of undoubted bovine 
origin. As, however, it is obvious that cow-pox was the 
source of the lymph stocks first introduced into use by 
Jenner and his contemporaries, it will be of interest to 
study briefly the nature and clinical appearances of this 
disease as seen in the cow and also in man ; whether 
accidentally contracted or intentionally inoculated. 

Cow-pox in the Cozv. — For a description of cow-pox 
in typical form, as it was known to Jenner and his con- 
temporaries, it is necessary to consult the writings of the 
early part of the century, at which period the attention 
of the medical and scientific world had been specially 


directed to this affection of cows by the teaching of the 
apostle of vaccination. Probably the most trustworthy 
accounts are those published by Bryce of Edinburgh, 
and later by Ceely ; and it is from their statements that 
the following description of the malady is derived. 

According to these observers, this affection, when 
once set agoing in a herd, tends to spread with con- 
siderable rapidity, the "matter" of the vesicles being 
carried by the milkers from one cow to another. It 
makes its appearance especially in the spring season, 
and is observed upon the udders and teats of the cows ; 
at first in the form of small vesicles containing a limpid 
fluid. These vesicles are of a bluish or livid colour, 
and are surrounded with considerable erysipelatoid 
swelling and inflammation. If ruptured, the vesicles 
tend to become irregular about the edges, and unless 
care be then taken are apt to degenerate into foul and 
troublesome sores. During the course of the affection 
the cow is not unfrequently observed to be in bad 
health; the appetite is impaired, the temperature is 
above normal, and the secretion of milk may be con- 
siderably diminished. If the material from the vesicles 
on the udders or teats of the cows happen to come in 
contact with an abrasion of the skin of the milker's 
hand, such person is apt to become infected with the 
disease. When the ailment is communicated in this 
manner, it is termed casual coiv-pox to distinguish it from 
that form which is intentionally propagated by inocula- 
tion, under which conditions the affection is less virulent 
than when communicated in the former way. Probably 


the more severe form in which casual cow-pox usually 
appears is to some extent due to the situation of the 
resulting vesicles, and to the purulent nature of the 
secretion from the sores on the cows' teats or udders. 

Casual Cow-pox in Man. — When cow-pox has been 
communicated to the milkers in the casual way, small 
inflamed spots appear in a few days upon the hands, 
more particularly about the joints and tips of the fingers. 
These spots quickly assume the appearance of small 
bHsters, somewhat resembling those from burns, which 
go on increasing until they become large vesicles of a 
circular form, with a flat or rather a concave surface ; 
their edges being considerably elevated above their 
centre. They have then acquired a somewhat bluish 
colour, and are found to contain a Umpid fluid. After 
some days the parts around the base of these vesicles 
become considerably swollen, hard, and inflamed, and, 
as the affection advances, they may assume something 
of an erysipelatous appearance. Pain and some degree 
of swelling of the axillary glands now denote an absorp- 
tion by way of the lymphatics, and, with the usual 
symptoms of fever, mark a constitutional affection which 
is sometimes so severe as to incapacitate the person 
from following his usual employment for some days. 
It does" not appear, however, that a general eruption 
ever follows even on the smartest attack of casual cow- 
pox. After a few days the pain, inflammation, and 
hardness of the surrounding parts gradually abate ; but 
the vesicles not infrequently ulcerate instead of becoming 
encrusted and drying up. These ulcerations, however. 

38 VA CCINA TION : chap. 

gradually heal up in course of time without occasion- 
ing any lasting injury ; and the constitutional affection, 
although severe, is always transient and unattended with 
danger : there is no case on record in which casual 
cow-pox is known to have been fatal. 

Inoculated Cow-pox in Man. — In the cow-pox induced 
by inoculation the appearances which present them- 
selves may differ considerably in some respects from 
those which have been described as occurring in the 
casual disease. 

Thus about the third day after the insertion of the 
virus of cow-pox, either by puncture or by slight incision 
in the arm, a small inflamed spot may be observed at 
the point where the inoculation was performed. 

Next day this spot appears still more florid, and on 
passing the point of the finger over it, a certain degree 
of hardness and swelling is readily perceptible. 

By the fifth day a small pale vesicle occupies the 
spot where the inflammation began, and the affection 
begins to assume the characteristic appearance of cow- 
pox. The vesicle has now a milky-white colour without 
any inflammatory zone around it, it is evidently 
depressed in the centre, and its edges are considerably 

For the next two days the vesicle increases in size 
and retains the same character, so that by the seventh 
day it has acquired very considerable magnitude ; if the 
inoculation be performed by a puncture, it assumes a 
circular form, if done by an incision, an oblong form. 
But in both cases the margin is regular and well 


defined, while the centre becoming still more depressed 
and the edges more turgid, the whole puts on an 
appearance which is very characteristic of this particular 

About the eighth day from the time of inoculation 
an inflammatory zone begins to appear round the base 
of the vesicle. This increases for two' or perhaps three 
days more, by which time it may be two inches or 
longer in diameter, and of a bright red colour. At this 
period, also, the vesicle still retains its concave appear- 
ance ; the crust in the centre has considerably increased 
in size, and begins to assume a dark or brownish 

About the eleventh day the vesicle has attained its 
greatest magnitude, and the surrounding inflammation 
begins to abate. The fluid in the vesicle, which before 
was thin and transparent, is now more viscid and 
slightly turbid. After this period the whole becomes 
quickly converted into a smooth, shining, and some- 
what translucent dry crust of a dark brownish or red 

This crust, unless forcibly removed, will adhere for a 
week or more, and then fall off, leaving the skin beneath 
apparently sound, but livid for a time, and more or less 
permanently scarred. 

In children little else than the above local process is 
usually noticeable, but in adults constitutional symptoms 
are apt to be somewhat severe. About the eighth day 
from the time of inoculation the glands in the axilla 
become a little swollen, and there is pain and stiffness 

40 VACCINATION chap, i 

on moving the arm. Headache, shivering, a rapid 
pulse, and other febrile symptoms present themselves, 
and these may persist for a period varying from a few 
hours to two or more days. 


Cow-Pox: I. Bryce. Inoculalion of Cow-Pox. Edinburgh, 
1802, and 2nd edition 1809. — 2. Ceely. Trans. Prov. Med. and 
Surg. Assoc, vol. viii. pp. 299-312, 342-352. — 3. Hering. Ueber 
Kiihpocken an Kiihen, p. 9, — 4. Lev. Experiments on the Origin 
of the Cow -Pox. Whitby, 1801. — 5. Rapport sur les vaccin. 
pratiqtdes eft France pendant 1841. — 6. Sixth Report of Medical 
Officer to the Privy Council, 1863, p. 10. 



Although it is clear from Jenner's writings that he 
believed vaccinia to be nothing more nor less than 
"smallpox of the cow," his theory appears not to have 
received general support even in his own day, and from 
that time onwards the value of the^ practice of vaccina- 
tion has by some been impugned on the plea that 
inoculation of one disease — " cow-pox " — could not be 
expected to exert any really protective influence against 
the ravages of smallpox, a disease supposed by them to 
be of totally different origin. ' And if the thesis of 
essential difference between these maladies were capable 
of demonstration, no doubt the objection would be of 
considerable weight. For there exists but little well- 
authenticated evidence that the living virus of one 
disease is capable, when inoculated into an animal, of 
affording protection against the effects of inoculation of 
the virus of another and totally different disease, although, 
no doubt, when two different viruses are inoculated at 
nearly one and the same time, the incubation period of 
one or other of them may be modified as to duration. 
In his first paper, Jenner, as has been said, advanced 


the thesis that smallpox and cow-pox are identical ; but 
even at the present day controversy wages hotly around 
this question, and it still awaits a definite solution. 
There can, however, be no objection to our speaking of 
vaccinia as "one with smallpox," since it matters Uttle 
from our present point of view whether smallpox, on 
its transference from man to the bovine animal, becomes 
actually transformed, or, as some would maintain, 
merely modified. During the long period which has 
now elapsed since the introduction of vaccination, 
many observers have set themselves the task of 
attempting, by experimental methods, to solve the 
problem of the true relationship of variola to vaccinia. 
These attempts have all been directed to the possibility 
of giving rise to cow-pox by the introduction in one 
or another manner of the virus of smallpox into the 
system of the bovine animal. In the great majority of 
such attempts, which are vastly more numerous than is 
generally supposed, the results have been entirely negative, 
although so numerous have been the experimenters, who 
from time to time have attacked the problem, that the 
total number of instances in which an apparently suc- 
cessful result has been obtained is now considerable. 

So far as I am aware, the first recorded experiments 
are those of Gassner of Gunzburg, in 1801, who suc- 
ceeded, after no less than ten fruitless attempts, in 
directly inoculating a cow with smallpox virus. The 
lymph thus obtained was employed for the vaccination 
of four children, from whom other seventeen were 
subsequently vaccinated. None of these exhibited any 


signs of smallpox, Viborg of Copenhagen, in 1807, is 
stated to have also been successful in variolating the 
cow. ^ Basil Thiele of Kasan twice, in 1836 and 1838 
respectively, succeeded in inoculating the cow with 
smallpox matter, and in vaccinating children with the 
lymph thus obtained. In the light of recent work on 
this subject, his conclusions are of much interest. As 
stated by himself, they are : |'(i) The so-called vaccine 
disease is not an eruptive dis'ease peculiar to the cow, 
but it is produced in it by the transmission of human 
smallpox to it, and the man and not the cow, as has 
hitherto been thought, is the source of the disease. 
(2) The mild disease thus caused in the cow can, by 
direct transmission from the cow to man, produce in 
him as mild a disease, which gives protection against 
the natural smallpox."] Thiele also described a method 
^y which "he 'clEifned^ to have brought about such modi- 
fication of smallpox virus as to render benign its action 
on the human subject, without the intermediate stage of 
implantation on the tissues of the cow. This altered 
potency of the material was brought about, according 
to his statement, by keeping the smallpox lymph for a 
period of ten days between two slips of glass, after which 
milk was added to it, and the resulting mixture employed 
for the vaccination of children. The same process was 
carried through with the lymph obtained from the first 
and following series of children vaccinated, until after 
ten generations addition of milk was found to be no 
longer necessary. 

Seeing that at this date (1836-38) true cow-pox was 


SO much more widespread than at present that at the 
commencement of the century Jenner confidentially 
looked forward to its occurrence every year, and that in 
consequence the majority of milch-cows were Hable to 
become infected at one or another period of their exist- 
ence, this empirical method of Thiele's bears a curious 
and noteworthy resemblance to one of the methods 
proposed by Koch for the treatment of rinderpest. 
This consists in the injection into susceptible animals 
of a mixture composed of a small proportion of virulent 
blood diluted with a much larger quantity of serum 
obtained from an immune animal. Milk, of course, is 
merely an emulsion due to suspension of fatty particles 
in a fluid which may be regarded as blood serum ; so 
that it is conceivable that milk obtained from a cow 
which had suffered not long before from cow-pox, and 
so become, for the time at any rate, immune, might so far 
modify smallpox lymph added to it, as to produce 
some such effect as noted by Thiele when the mixture 
was used for purposes of inoculation. I have been 
unable to find any records dealing with a repetition of 
these experiments, but it would be interesting to make 
investigation of the possibility of producing a similar 
lessening in virulence of smallpox lymph by admixture 
with the blood-serum of a calf which had been vaccinated 
some few weeks beforehand. 

Ceely, in February 1839, inoculated three stirks 
with variolous lymph, with the result that in two of the 
three he eventually obtained, in due course, what he 
considered to be vaccine vesicles. From these, lymph 


Stocks were established, which at once came into such 
extensive use that in the course of a few months at least 
a couple of thousand children had been vaccinated 
therefrom. In his paper recording these experiments, 
he states that on former occasions, even though working 
purposely at different times of the year, he had altogether 
failed in attempts to variolate the cow.' In order to 
test the measure of protection afforded to these children 
by his variola-descended lymph, Ceely tried on a number 
of them the effect of subsequent variolous inoculation 
at varying intervals of time from their vaccination. In 
no case did he obtain any greater result locally than, 
on the authority of Willan, is known to have attended 
the test-inoculations employed by Jenner and Woodville. 

Unfortunately, owing to the particular methods used 
by Ceely in his variolous inoculations of cows, some 
doubt is thrown on the value of the results obtained by 
him. He was, according to his own account, working 
simultaneously with both variolous and vaccine lymph, 
so that it is difficult to exclude the possibility that his 
instruments may have unintentionally served to convey 
the infection of both diseases at one and the same time. 

Several years before the commencement of his 
inoculation experiments, Ceely had made trial of a 
method originally employed by Sonderland of Barmen 
(1830), who claimed to have infected cows with the 
contagion of variola, by way, presumably, of the 
respiratory tract. This he did by enveloping them in 
blankets taken from the bed in which a patient had 
died of smallpox, and by hanging other blankets 

46 VA CCINA TION : chap. 

similarly infected around the head of each cow, in order 
that the animal might breathe in the effluvium arising 
therefrom. It is stated that these cows subsequently 
manifested symptoms of cow-pox, and that from this 
source lymph was obtained and used on the human 
subject with the result that typical vaccine vesicles were 
produced. Ceely failed, however, in his attempt to 
bring about infection in similar fashion, as, indeed, have 
numerous observers in various parts of the world. Yet 
Ceely has himself recorded an occurrence observed by 
him in 1840, which although apparently but little known 
is of interest in this connection. He relates an instance 
in which five out of eight milch cows sickened with 
cow-pox within twelve and fourteen days of their having 
been seen to be licking over a quantity of flock from 
the mattress on which a patient had died of confluent 
smallpox, and which had been spread out in the field 
for purification. Careful inquiry at the time appears to 
leave no doubt that the animals, which had been on the 
same farm for a considerable period, were in good health 
at the time of their admission to the meadow in which 
the bedding was exposed, and also that there had not 
been any other case of cow-pox in the neighbourhood. 
The fact that all the animals became affected simultane- 
ously, and after a period corresponding to the usual 
incubation period of variola, certainly affords some 
reason for the assumption that the outbreak owed its 
origin to the animals being exposed to the " effluvium " 
from the infected bedding. It is of course possible that 
infection by way of the digestive tract may have been 


a factor also in this case, since Chauveau and other 
observers have shown experimentally that such result 
may be possible, a number of cases having now been 
recorded in which vaccinia has been communicated both 
to the human subject and also to the lower animals by 
the ingestion of vaccine lymph or powdered vaccine crusts. 
There surely has seldom been so indefatigable a 
repeater of experiments as Badcock the Brighton chemist, 
who during a period of about twenty-five years per- 
formed more than five hundred variolous inoculations. 
In December 1840, Badcock, who had commenced his 
work without knowledge of Ceely's slightly earlier work, 
first succeeded in variolating a cow, from which he 
carried on a stock of what was indistinguishable from 
genuine vaccine lymph. Although he subsequently 
met with similar success on no less than thirty-seven 
separate occasions, the difficulty and uncertainty of the 
work will be appreciated on consideration of the fact 
that those experiments, which eventuated in success, 
represent but 7 per cent of the total number. The 
lymph obtained by Badcock as the result of his variolation 
experiments has been largely employed \ he supplied it 
to hundreds of medical practitioners, and many thousands 
of children are said to have been successfully vaccinated 
with it. About twelve years (1852) after Badcock had 
obtained his first successful result, two American physi 
cians, Drs. Adams and Putnam, were equally fortunate, 
and were able in consequence, as reported at the time in 
a Boston daily paper, to furnish all the vaccine matter 
requisite for use in the city and neighbourhood. 

48 VACCINATION: chap. 

In 1865 a Commission, appointed by the Societji of 
Medical Sciences at Lyons, having Chauveau as its 
head, reported the results of a comprehensive series of 
experiments carried out by it during the two preceding 
years, the results of which were completely at variance 
with those obtained by previous investigators. These 
results have been admirably summarised in the Report 
of the English Royal Commission on Vaccination, 
" Inoculation of the cow with smallpox matter in 
any one of the thirty animals used did not give rise 
to a vaccine vesicle ; nevertheless, a definite result was 
obtained in the form, however, not of a vesicle, but of 
a thickening and inflammation of the wound ; when a 
puncture had been made this became a papule ; lymph 
squeezed from such a papule, and inserted into a second 
animal, gave rise to a like papule ; and this again might 
be used for a third animal, but often failed; and the 
effect could in no case be carried on through more than 
three or four removes. When the inoculation was re- 
peated on an animal on which a previous inoculation 
had produced such a papule, no distinct papule was 
formed; and, moreover, lymph squeezed from the seat 
of the latter inoculation produced no effect at all when 
used for the subsequent inoculation of another animal." 
There is evidence that the development of the papule 
was the result of the specific action of the virus. This 
inference is strengthened by the fact that no such papule 
was produced by the Lyons Commission when the 
smallpox matter was inserted into an animal which had 
previously had cow-pox naturally or artificially ; as well 


as by the fact that when an attempt was made to 
vaccinate, with vaccine lymph of proved efficacy, an 
animal on which a papule had been so developed by 
inoculation with smallpox matter, the vaccination failed, 
though the animal had never had natural cow-pox nor 
been vaccinated. The specific nature of the lymph of 
the " Lyons " papule is held to be shown by the fact 
that such lymph, when used on the human subject, gave 
rise to smallpox. On the other hand, it has been 
urged that in this case ■ the virus producing the effect 
was simply the original smallpox matter used in the 
inoculation, producing the papule and still clinging to 
the wound. This, however, is considered to be dis- 
proved by the experience that lymph from a " Lyons " 
papule of the second remove also gave rise in the human 
subject to smallpox. Thus Chauveau and his Com- 
mission found that smallpox implanted in the bovine 
animal gave rise to a specific effect which was not cow- 
pox, but was of the nature of smallpox ; though its 
manifestations in the cow were different from those of 
smallpox in man. 

A few years earlier, in i860, Martin of Boston, U.S.A., 
appears to have had a somewhat similar experience, as 
about fifty persons vaccinated by him from vesicles 
raised on a cow's udder by inoculation of variolous 
matter taken from a pock on the body of a man who 
had died of the disorder, were nearly all attacked with 
smallpox, and three died. 

Voigt, the Director of the Hamburg Vaccine 
Establishment, in 188 1, brought into extensive use not 


50 VACCINATION : chap. 

only in Hamburg, but also in other parts of Germany, 
a strain of lymph of variolous origin. In the early 
summer of that year he inoculated three calves with 
lymph obtained from a case of smallpox. In two of these 
the operation was apparently unsuccessful, in view of 
which it is of interest to note the fact that all three calves 
were afterwards found to be insusceptible to vaccina- 
tion. In the third calf, a single vesicle appeared on the 
site of one, out of five incisions made on the left side 
of the perineum. Voigt adds, however, that this same 
animal had been vaccinated in one small place in the 
right hypochondrium (for what reason is not obvious), 
and that at this point a normal vesicle developed. 
Voigt does not consider that this fact invalidates the 
apparent success of his experiment, for the reason that, 
as he says, the lymph derived from, as he considers, 
the variolous source proved to be more potent, as 
regards its observed effects on children and calves, than 
the Beaugency vaccine lymph, which he had in current 
use at the time his experiment was carried out. Both 
Chauveau and Berthel have, on the other hand, 
expressed the opinion that Voigt's variolo- vaccine is 
merely a continuation of the strain of animal vaccine, 
previously employed in Hamburg. 

In 1885 Simpson succeeded in an attempt to inoculate 
a cow with smallpox lymph obtained from a young 
vaccinated girl, but, so far as I am aware, he did not 
publish the details of this experiment at the time. Dr. 
Cory, to whom Simpson sent points charged from the 
cow on the sixth day after inoculation, has recorded the 


results following on their use in his recent volume on 
Vaccination. Three days after receiving the points, Dr. 
Cory employed them at the Government Animal Vaccine 
Establishment, Lamb's Conduit Street, for the vaccination 
of a young male calf. Three out of five insertions made 
with the lymph resulted in vesicles indistinguishable 
from those produced by the current lymph of the 
Establishment. A second calf and also a child were 
therefore vaccinated from the first calf, with successful 
results in each instance, although in the calf only six 
vesicles resulted from seventeen insertions of the lymph. 
From this calf a continuous series of calf vaccinations 
were carried on during a period of six months. With 
the lymph obtained from these animals, 79 in number, 
no less than 1247 children were vaccinated, 11 74 of 
whom afforded an insertion success-rate of 98.4 per cent. 

Some years later (in 1892) Simpson, then resident 
in Calcutta, was again successful in raising a stock of 
vaccine lymph of variolous origin. Lymph was taken 
from a smallpox patient on the fifth, sixth, seventh, 
and eighth days of eruption, and each day a young calf 
was inoculated, the animal having been taken into the 
smallpox ward for that purpose. 

The inoculations with the three later lymphs were 
unsuccessful, but the first calf showed on the sixth day 
of inoculation three papules on the abdomen and in 
the groin, at points where no punctures or incisions had 
been made. These on the following day had developed 
into vesicles. Two of the incisions and one puncture 
also contained lymph. On the sixth day lymph was 

52 VACCINATION : chap. 

taken (whether from vesicles, incisions, or puncture, or 
all three is not clear from Dr. Simpson's paper) for the 
inoculation of another calf, from which again, on the 
sixth day, a child was vaccinated, the two insertions 
giving rise in due course to vesicles. From this child 
the strain of lymph was carried on successfully through 
nine generations, and Dr. Simpson remarks that each 
child showed splendid vaccine vesicles, much more 
typical in every respect than those obtained from the 
lymph, which had been previously in use. 

In 1886 and 1890, Fischer, the Director of the 
Institute for Vaccination at Carlsruhe, obtained two 
strains of variolo-vaccine, as the result of inoculation of 
calves with human smallpox lymph. In the first series, 
Fischer inoculated a young female calf, which he took 
into the smallpox ward of the Pforzheim Hospital for 
this purpose. Typical vaccine vesicles had developed 
on the sixth day, the lymph from which was carried on 
through twelve generations in the calf, after which it 
was successfully employed for the vaccination of 
children. Fischer states that the initial experiment of 
his second series was carried out in the Carlsruhe 
Institute, but that the animal was placed in a separate 
stable, and that every precaution was adopted to secure 
freedom from vaccine contaminations. In this calf, 
directly inoculated with smallpox lymph, he again 
obtained, as in his experiments of four years previously, 
a vesicular eruption, having the characteristic appearance 
of ordinary vaccinia. 

With lymph taken at the third remove from this 


calf, he vaccinated his own grandson with complete 
success, and the strain, which has now been continued 
through a very large number of animals, has been 
brought by Fischer into general use. 

Surgeon-Major King in 1889, being then stationed 
at Madras, inoculated a young bull-calf with fifth day 
smallpox lymph obtained from a child suffering from 
the disease. On the eighth day after the inoculation, 
a crop of vesicles made their appearance, not at points 
of insertion, but some little distance away, from the lymph 
obtained from which another calf was then inoculated. 
In this calf again secondary vesicles appeared, but, in 
addition, vesicles also formed at each point of insertion 
of the lymph. The lymph stock was carried on through 
three more generations in calves, the vesicular eruption 
being now in each instance strictly localised to the 
points of insertion. With lymph obtained from the 
fifth calf of the series, a number of children were 
vaccinated successfully. Dr. Simpson reports that 
from the stock thus established by King, 4240 British 
and native soldiers have been vaccinated, and also 
a very large number of the native civil population, 
with most gratifying results ; the percentages of success 
being much above that obtained from the lymph 
previously in use. For reasons which are unknown to 
me. King's investigation appears to have been regarded 
with disfavour by the Government of Madras, by whom 
he was ordered back to his regiment and deprived of 
his special emoluments. Subsequently, however, this 
injustice was repaired, by the reinstatement of Surgeon- 


Major King in his former position as Director of 
Vaccination in the Presidency. 

In May 1892, Hime pubhshed the results obtained 
by him in a single experiment on variolation. The 
lymph employed for inoculation of the first calf was 
collected from a semi-confluent case of smallpox in a 
previously vaccinated woman of thirty-seven years old. 
No details are given as to tvhere the experiment was 
conducted ; but presumably it was the private establish- 
ment from which Dr. Hime supplies vaccine lymph, as 
he makes the statement that " the calf was placed on 
the ordinary vaccination table used for these animals." 
A number of skin incisions were made, into which the 
variolous lymph was inserted. Of these incisions all 
subsequently healed, being on the sixth day only 
" detectable to the touch by a slight roughness." Several 
"pocks," however, developed at points for the most 
part distinct from any of the incisions, and material 
taken from these on the ninth day of inoculation was 
employed for the direct inoculation of another calf. 
All the incisions made on this second calf had " taken " 
on the fifth day, when lymph was collected in tubes, 
and pulp removed by scraping. Some of the latter 
made into " conserve " with glycerine was sent to 
Fischer of Carlsruhe, who obtained excellent results 
with it on a calf, from which children were in turn 
successfully vaccinated. Hime also himself vaccinated 
a child with a capillary tube of the lymph in two inser- 
tions, at each of which points a typical vesicle presented 
itself on the eighth day. 


The largest amount of research work in this direc- 
tion during recent years is that of Haccius and Eternod 
(1893), of whom the former is the Director of the Swiss 
Vaccinal Institute at Lancy, Geneva, where the work 
w^s carried out. Haccius gives an account of no less 
than seven successful series of experiments, in each of 
which a strain of variolous lymph, obtained in the first 
instance from the human being, was carried on from 
calf to calf through, in some instances, as many as six or 
seven removes. The general result was that by the 
second, or at the most the third, remove, the effect 
produced was altogether indistinguishable from that 
which we are accustomed to see in a calf that has been 
successfully vaccinated. With the lymph of the fifth or 
sixth remove, a number of previously unprotected 
children were vaccinated with unfaihng success. The 
resulting vesicles could in no way be distinguished from 
ordinary vaccination vesicles of equal age. The total 
number of animals employed in these experiments is 
not mentioned, but from certain statements in M. 
Haccius' publication, one gathers that it was very con- 
siderable. It is obvious, therefore, that the Lancy 
experience, as to the small proportion of success to 
failures, is similar to that of other observers. It is 
worthy of note, also, that in those animals in which 
inoculation of variolous matter appears to have been 
attended with success, no definite vesiculation was ever 
observed at the site of any of the punctures or incisions 
that were made. The usual result in the first remove 
from the human being was either a single vesicle or 

56 VACCINATION : chap. 

group of vesicular points on the site of a scarifica- 
tion or denudation of the skin. It is particularly 
stated that every precaution was taken, as, for in- 
stance, by the sterilisation of the lancet before each 
operation, to prevent the accidental transmission of 

Klein, who in 1879 had obtained in thirty-one trials 
what then appeared mere negative results, renewed his 
investigations in 1892, employing now calves for inocu- 
lation experiments instead of milch cows, as in his 
earlier attempts. Having procured at the Hospital Ships, 
off Dartford, lymph from the vesicles of two cases of 
confluent smallpox, one at the sixth, the other at the 
seventh day after appearance of the rash, this lymph 
was, on the same day, inoculated by linear cutaneous 
incisions on two calves at the Brown Institution. The 
inoculation of one of these calves afforded no indication 
of success, but on the other calf he obtained a local 
result in the groin, which, however, consisted not of a 
distinct vesicle, but merely a thickening and redness, 
together with a linear crust, at the site of certain of the 
incisions into which smallpox lymph had been intro- 
duced. Lymph pressed from the thickened wounds, 
when inoculated into a second calf, produced also by 
the fifth day a like but rather more marked result ; 
while the thickening and redness still further increased 
as the process was repeated in a third and fourth calf. 
In none of these animals was there any appearance of 
vesiculation. At this stage the fourth calf of the series 
was removed to Lamb's Conduit Street, where Dr. 


Cory, with material obtained by clamping and scraping 
the thickened incisions on the seventh day, vaccinated 
an infant in five separate insertions, with complete 
success. Each of these presented on the eighth day 
appearances characteristic of vaccinia. Moreover, Dr. 
Cory, who kept the child under observation from first 
to last, was unable to detect in the course of the dis- 
order any deviation from the condition of affairs normal 
to vaccinia. " Crusts " from the arm of this child were 
preserved and utilised, after being mashed up in sterile 
salt solution for the retro-vaccination of another calf. 
In due course there resulted, in about half of the total 
number of incisions, vesiculation of a character not to 
be distinguished from that current in the calves vacci- 
nated in the ordinary course at the Lamb's Conduit 
Street Station. Finally, after a lapse of six weeks, 
Klein submitted this retro-vaccinated calf and other 
calves vaccinated from it to the test of a further and 
thorough vaccination with current calf lymph, and with 
the result that all these animals entirely resisted vaccinia. 
I have learned privately from Dr. Klein that, in his 
former (1879) experiments on cows, he in more than 
one instance obtained results somewhat similar to those 
witnessed in the calf on which his more recent success- 
ful inoculation was performed. He suggested at the 
time the advisability of attempting a transference to a 
second animal, but Ceely, who was superintending the 
experiments, insisted on the necessity of obtaining a 
vesicle in the first animal inoculated as the only 
criterion of success, and the suggestion was therefore 


overruled. For his own satisfaction, however, Klein 
endeavoured to vaccinate one of the cows which had 
shown some local reaction, but entirely without result, 
the animal apparently being immune. 

Of my own experiments the most successful series 
may briefly be described as follows : — With variolous 
lymph (contained in two capillary tubes) taken on 30th 
July 1892 from a vaccinated girl, aged sixteen years, at 
from the fifth to the sixth day of the eruption of discrete 
smallpox, I inoculated on nth August a cow-calf A, 
about six weeks old, by thirty-two linear incisions and 
two superficial scarified patches made on the abdomen. 
On 13th August (the third day) this calf presented 
nothing but a slight scab over the scarified patch. On 
15th August (the fifth day) some of the incisions were 
somewhat red and elevated, more particularly at several 
definite points which tended to be vesicular. At this 
date, on the inner aspect of each thigh, and distant in 
each instance from incisions or scarifications, was a crop 
of shotty and incipiently vesicular pimples. On 17th 
August (the seventh day), having clamped three of the 
incisions, including those with the quasi - vesicular 
appearance, I removed the lymph with the crusts by 
scraping, and used the material for the inoculation on 
this date of calf B. The accessory incipient vesicles I 
left untouched. From first to last calf A never exhibited 
any appreciable rise of temperature or indisposition of 
any sort. On the ninth day the linear incisions, the 
cross scarifications, and the accessory vesicles of calf A 
were in their decadence, and in a few days more the 


whole process was at an end. More than a month 
later, namely, on 22 nd September, I vaccinated this 
calf A in twenty-three linear incisions, with calf lymph 
obtained from the Animal Vaccine Establishment. The 
result was absolutely negative. 

On 17th August, with the scrapings obtained the 
same day from calf A, I inoculated a cow-calf B, two 
months old, in fifty-six linear incisions on the abdomen. 
On the third day every one of these incisions was 
distinctly raised and bordered by a deHcate pink flush. 
On the fourth day (20th August) all incisions had 
evidently "taken," the areola about each being now well 
marked, and about one-eighth of an inch in width, but 
there was no definite vesiculation. At this date, after 
clamping, scrapings were taken from certain of the 
incisions for further experiment. No accessory vesicles 
appeared in this calf, which, along with calf A, was 
vaccinated on 22nd September in seventeen linear 
incisions with calf lymph obtained from the Animal 
Vaccine Establishment. The result, as in calf A, was 
absolutely negative. 

On 20th August, with scrapings taken the same day 
from calf B, I inoculated a small cow-calf C (7 weeks 
old) in twenty-seven linear incisions on the abdomen. 
On the fourth day (23rd August) every incision had 
"taken," and in most of them there was evidence of 
commencing vesiculation. Two days later (25th August) 
vesiculation was distinct in several of the incisions. At 
no time were there any accessory vesicles. This calf 
also was vaccinated (22nd September) with calf lymph 

6o VACCINATION : chap. 

from the Animal Vaccine Establishment. Again the 
result was altogether negative. 

I have thus far obtained an undoubtedly successful 
result in one series only out of four attempts, but I have 
at any rate been able to satisfy myself that it is possible 
to variolate the calf, and, further, that the result obtained 
in the first instance may become greatly modified in the 
course of successive removes ; and, again, that animals 
which have been thus treated are no longer susceptible 
to vaccination. 

I may perhaps mention here that all my experiments 
were conducted at the Brown Institution, to avoid any 
possibility of contamination with vaccinia ; that, as a 
further precaution, new scalpels were used, which were 
invariably first carefully sterilised in a flame ; that before 
and after use the table was thoroughly washed with 
carbolic acid and hot water, and during the intervals of 
use kept exposed to the open air. 

The Royal Commission on Vaccination, in the section 
of their report dealing with the question of the relation- 
ship of variola and vaccinia, show that the various series 
of inoculation experiments, of which I have given a brief 
account, fall into one or other of three categories. The 
first category includes the experiments of Thiele, Ceely, 
Badcock, Voigt, Haccius and Eternod, King, Simpson, 
Hime, and others. In these experiments " inoculation 
of smallpox matter into the udder, or adjoining parts, 
of the bovine animal gave rise, at or near the site of 
inoculation, to a vesicle, either identical in visible 
characters with the ordinary vaccine vesicle produced 


by inoculation with the matter of cow-pox, or to a 
vesicle the features of which, while not corresponding 
wholly with those of a perfect vaccine vesicle, so closely 
resembled it as to justify the recognition of the vesicle 
as a vaccine vesicle. Also it includes experiments in 
which, though the local result had not the characters of 
a perfect vaccine vesicle, yet lymph from it, when 
carried through a second or third remove in the cow or 
calf, presented results fully manifesting those characters, 
and when again transferred to man gave results in- 
distinguishable from the ordinary vaccine vesicles." In 
the second category are placed the experiments of Klein 
and myself. In no instance did either of us obtain any 
appearance of vesiculation in the animals directly in- 
oculated with smallpox virus ; although the appearance 
of the eruption tended to resemble more and more that 
typical of vaccinia, the further the remove, through a 
series of calves, from the original strain. In the third 
category are included the practically negative results of 
the Lyons Commission, obtained by Chauveau and his 
fellow workers. A brief account of these has already 
been given. 

With the exception then of Martin and of Chauveau 
and his colleagues of the Lyons Commission, all the 
observers mentioned claim to have obtained positive 
results, in a certain number of their experiments, at any 
rate, as regards the production of typical vaccinia, after 
one or more removes, as the result of variolation of the 
cow or calf. By no one, apparently, has success been 
attained invariably ; but it is among the experiments of 


the earlier observers especially, who made use, for the 
most part, of heifers and milch cows, that the largest 
proportion of abortive attempts are to be met with. 
Subsequent experience has shown that success is much 
more likely to be attained if calves be used instead of 
heifers or cows. In this way, perhaps, Chaveau's some- 
what anomalous results may be in part explained. 

With reference to recent experiments on variolation 
of the calf, it is worthy of note that, as previously 
mentioned, different observers have obtained local 
effects in that animal which, in different calves of a 
series, have varied considerably. The final result has, 
however, after a greater or less number of removes from 
calf to calf, been invariably the same ; namely, a local 
vesicle is produced which by no means at our command, 
such as the appearance and course thereof, or the pro- 
tective power of the lymph derived therefrom, is dis- 
tinguishable from true vaccinia. 

Although practically there is unanimity of opinion 
among those who have worked at this subject, it must 
be confessed that, seeing the conditions under which 
they were carried out, many, particularly of the earlier 
experiments, are of little worth. Some of the main 
objections are based on the fact that experiments on 
this subject have almost without exception been per- 
formed in estabhshments devoted to the continuous 
cultivation of vaccinia; on the use concomitantly of 
vaccine and of variolous lymph on the same animal ; 
and on the want of care as to the cleanliness and 
freedom from vaccine contamination of lancets and 


" points " used in the experiments. Objection of 
similar sort against the variolations of the calf, which 
have been achieved in recent years, is hardly valid. In 
some of the more recent cases, at least, special pre- 
cautions have been taken to ensure that the instruments 
and table were sterilised, and to render the environment 
of the animal such as to afford no likelihood of the 
communication of vaccinia. Most especially was this 
so as regards Klein's investigations and my own. The 
number of successful cases which have been recorded is 
now so large that it is difficult to believe that sources 
of fallacy of the above sort should have been present in 
every instance ; and it is therefore well-nigh impossible 
to resist the conclusion that a change of smallpox into 
vaccinia must really have come about. 

Nevertheless, there are found even at the present day 
those who, like Juhel and Dupuy (1894) and Layet 
(1895), maintain the essential duality of variola and 
vaccinia. It is, however, difficult to understand how 
the results obtained, by the last -mentioned especially, 
can bear the interpretation he would place upon them, 
seeing that he obtained, as the result of inoculating 
heifers with smallpox lymph, a vesicular eruption, the 
lymph derived from which reproduced the disease on 
other heifers. Moreover, subsequent vaccination of all 
his series of animals was either abortive or wholly with- 
out result. 

Hervieux(i895), the Director of the Vaccine Institute 
attached to the Academic de Medecine at Paris, takes 
up a similar position to that of Layet and his collabo- 


rators, Le Dantec and Benech. He asserts that the only 
point of similarity between variola and vaccinia, which 
has been brought forward by those who believe in the 
relationship of these disorders*, is the likeness between 
the vesicles which are found in either case, and denies 
that this point forms any argument in favour of their 
identity. He also considers that the use of lymph of 
variolous origin is fraught with danger, and so, doubt- 
less, it may be when that employed is derived, as in 
Chauveau's and Martin's cases, for instance, directly 
from the animal first inoculated. 

If, as I believe, it can be conclusively proved that 
smallpox lymph, by passing through the system of the 
calf, can be so altered in character as to become deprived 
of its power of causing a generalised eruption, while 
inducing at the site of inoculation a vesicle in- 
distinguishable from a typical vaccine vesicle; and, 
more important still, if it be shown that when trans- 
ferred again to man, it has by such treatment completely 
lost its former power to produce a general disease, it 
may fairly be asserted that cow-pox — or rather that 
artificially inoculated form of the disease which we term 
vaccinia — is nothing more nor less than variola modified 
by transmission through the bovine animal. Perhaps the 
most reasonable interpretation of such results may be 
that smallpox and vaccinia are both of them descended 
from a common stock — from an ancestor, for instance, 
which resembled vaccinia far more than it resembled 
smallpox. It is conceivable, indeed, that the seeming 
vaccinia, obtained in the calf by inoculation of small- 


pox matter into that animal, may after all be but a 
reversion to an antecedent type ; and in this connection 
we may call to mind a fact of universal experience, 
namely, that vaccinia, however it may have arisen in the 
past, or is made to appear in the present, exhibits little 
tendency to " sport " (as, for instance, by manifesting 
a " generalised eruption ") in the direction of small- 

Mr. Picton and Dr. Collins, in their addendum to 
the Report of the Royal Commission on Vaccination, 
lay much stress on the want of " evidence to show that 
inoculation of the pox of the cow on the human skin 
has ever produced smallpox." Variola and vaccinia 
may, nevertheless, have a common ancestry, since it is 
not unlikely that variola may have departed widely from 
the original type, and have gained an exalted virulence 
by repeated passage through man under conditions 
favourable to its propagation and activity. If this 
evolution of the disease has, in fact, taken place, variola 
may have suddenly reverted, under greatly changed 
conditions (as for instance implantation on the bovine 
animal), to an ancestral type. But the reverse process 
ij.e. sudden " sport " of vaccinia in the direction of 
smallpox) is not to be expected. It is most unlikely 
that a less differentiated form (cow-pox), also emanating 
from the common ancestral stock, should attain to the 
most exalted virulence in a single individual, and per 
saltum declare itself as smallpox, as the dissentient 
Commissioners insist that it ought to do. 


Relationship of Variola and Vaccinia: i. Badcock. 

Experiments conjirniing the Power of Cow-pox, etc., 1 840. — 2. 
Ceely. Trans. Prov. Med. and Surg. Assoc, vol. viii. pp. 379- 
402, — 3. Chauveau, Viennois, and Meynet. {Rapport par) 
Md moires et coniptes-rendtis de la Soc. mM. de Lyon, tome v. — 4. 
CoFEMAN. Trans, of Epidemiological Society, 1 892-93 ; Journal 
of Pathology and Bacteriology, Mdiy \^^/^. — 5. Hime. Brit. Med. 
Journal, yo\. ii. p. 1 17; 1892. — 6. KiNG. Trans. South Indian 
Branch Brit. Med. Assoc, vol. iv. No. i ; 189). — 7. Klein. 
Report of Medical Officei- to the Local Government Board for 1891- 
92; (1893). — 8. Macpherson and Lamb. Trans. Med. and 
Phys. Soc. of Calcutta, vol. vi. and vol. viii. — 9. M 'Michael. 
Report of the Vacc. Sec. of Prov. Med. and Surg. Assoc. 1839, p. 
24. — 10. Freyer. Zeitschrift fiir HygieJie, Bd. xxiii. H. 2, 
1897, — II. Simpson. Indian Medical Gaz. May 1892, p. 148. — 
12. SONDERLAND. Hufcland's Journal, ]2Xi. 1831. — 13. Thiele. 
Henke's Zeitschrift, 1839, Heft i. 



Jenner's first case of vaccination was, as previously 
stated, that of a boy eight years of age, whom he 
inoculated in the arm with cow-pox matter taken from 
the sore on the hand of a dairymaid, who, in turn, had 
become infected with the disease from milking cows 
suffering from cow-pox. 

This was in 1796, and was his first actual experiment 
purposely performed, though it is evident that for years 
previously he had taken note of the results of the 
testing of many experiments unintentionally performed 
by milkers on their own persons, such experiments 
being subsequently tested at a longer or shorter interval 
by inoculation with smallpox, the inoculation being 
itself in many cases performed not purposely as a test, 
but as a preventive of smallpox in the course of the 
ordinary practice of some medical man in the dairy 
districts. In experiments and tests so conducted, there 
could be no " personal equation " to confuse the result. 

It was apparently not until 1798 that he made his 
first attempt to carry on a strain of lymph from arm to 
arm. In the spring of this year he inoculated a child 


with matter taken directly from the nipple of a cow ; 
and from the resulting vesicle on the arm of this child 
first operated on, he inoculated, or, as it may now be 
more correctly termed, "vaccinated" another. From 
this child several others were vaccinated ; from one of 
these a fourth remove was carried out successfully, and 
finally a fifth. Four of the children were subsequently 
inoculated with smallpox — the " variolous test " — with- 
out result. 

At this point, however, the strain appears to have 
been allowed to die out; but, in January 1799, Wood- 
ville, physician to the Smallpox Hospital in London, 
who had been much interested in Jenner's investigations, 
discovered the presence of cow-pox in a dairy in Gray's 
Inn Lane. With lymph taken, in the presence of Sir 
Joseph Banks, the President of the Royal Society, and 
other well-known men of the time, from one of the cows 
in this dairy, he vaccinated seven persons at the Small- 
pox Hospital ; while in the case of certain other persons, 
he employed matter from sores on the hand of a dairy- 
maid, who had become infected from one of the cows at 
this same place. These cases, from which afterwards in 
succession many hundreds of persons were vaccinated, 
were the main source of what is usually spoken of as 
"Woodville's lymph." 

These strains of lymph were extensively distributed 
by Woodville. Dr. Pearson, one of the surgeons to 
St. George's Hospital, also sent out much lymph. 

He very early obtained lymph from a dairy in the 
Marylebone Road. It is not unlikely also that a con- 


siderable part of his lymph was obtained from Wood- 
ville's cases. Jenner himself used some of Woodville's 
lymph, and he obtained a further supply from a cow at 
Mr. Clarke's farm in Kentish Town. 

The lymph first employed on the Continent and in 
other foreign countries was undoubtedly supplied in large 
measure by Pearson and Woodville ; although we learn 
from Baron and other authors that Jenner, who was 
naturally much appealed to for supplies of lymph, him- 
self sent lymph to Stromeyer of Hanover, to De Carro 
of Vienna, to Berlin, and to Barbados, Newfoundland, 
and other parts of America. 

The lymph which Woodville first sent to Paris 
died out, and he himself visited France with a fresh 
stock, taken at a time when he had learned to be 
careful to avoid contamination of his stock by variolous 

Strains derived from lymph stocks originally supplied 
by Jenner were also sent abroad by a number of 
different persons, the original strain being,, in large 
measure at any rate, the lymph obtained by Jenner from 
the Clarke's farm cow. Thus Ring records that he 
distributed Jenner's lymph to various places on the con- 
tinents of Europe and America. 

It would be erroneous to suppose, however, that 
all the lymph employed abroad in the early days of 
vaccination was obtained from England. Indeed, both 
Sacco and De Carro made extensive use of lymph 
obtained by the former from a case of natural cow- 
pox which he discovered in Lombardy. , In a letter 


to Jenner, under date i6th October 1801, Sacco states 
that from this source "more than eight thousand 
inoculations " had already been " performed with the 
most happy success," and he adds " several hundreds of 
these have since been subjected to the variolous inocula- 
tion, and have resisted it." In a postscript he adds that 
he had sent some of the Lombardy vaccine matter to 
Woodville. From this stock also De Carro sent supplies 
to Constantinople, where it was employed for the first 
vaccinations carried out in this part of Europe. De 
Carro it was also who first succeeded in conveying a 
supply of lymph to India. This lymph, again, was not 
from Jenner's stock, but was of Milanese origin, having 
been furnished to him by Sacco. It was, moreover, not 
of bovine but of equine origin, and according to De 
Carro had never been passed through the cow. 

This strain of equine lymph was originally obtained 
by Sacco in 181 2, from a crop of vesicles on the hands 
of a coachman who had not had smallpox, and who had 
dressed the heels of a horse affected with the "grease." 
From these vesicles two children were successfully 
vaccinated, and from these the stock was carried on 
through a long series of vaccinations. 

Among more recent strains may be mentioned that 
obtained in 1836 at Passy in the environs of Paris from 
the hand of a milker, who had contracted casual 

The old stock then in use at the Academic de 
Medecine had evidently degenerated somewhat ; and, 
when its effects were compared with those of the new 


Passy lymph, the vesicles developed from the latter were 
found to be manifestly finer. 

In 1836, Estlin of Bristol put in circulation a stock 
which at first showed unusual activity. This abated, 
however, after some transmissions, and the lymph after- 
wards came into extensive use. 

From this time onwards the various stocks became 
so numerous that Ceely, writing in 1841, states that 
during the preceding three years he had experimented 
with lymph from more than fifteen distinct sources ; of 
these six had been taken from the natural disease, either 
direct from cows or from vesicles on the hands of the 
milkers, and seven were artificially produced in the cow. 

The lymph stock in use at the present time at the 
Government Animal Vaccine Establishment was originally 
obtained on 26th November 1881 at a farm in the 
village of Laforet not far from Bordeaux; whence a 
sample of lymph from the seventeenth calf in succession 
from the animal first affected was sent by Dr. Dubreuilh 
of Bordeaux, to the Medical Officer of the Local Govern- 
ment Board. 

Of late years, more particularly, numerous strains of 
so-called variola-vaccine lymph, obtained by inoculation 
of human smallpox on the calf, have been introduced, 
especially by Fischer in Germany, by Haccius in 
Switzerland, and by King in India. These strains have 
been successfully transmitted through many thousands 
of individuals. 

Those who deny the relationship of smallpox and 
cow-pox will say that these children have been variolated, 


and not vaccinated. One, of course, admits that they 
have been variolated, in the sense that they have been 
inoculated with lymph descended from a case of human 
smallpox, but differing from the mild inoculations of 
Adams, Dimsdale, and the Suttons, in that the resulting 
disease is no longer infectious. Such procedure is also 
strictly comparable with those methods of protective 
inoculation by the use of attenuated virus, which of late 
years have given valuable results in the prevention and 
treatment of various zymotic diseases. 


Lymph Stocks: i. Baron. Life of Jenner^ vol. i. — 2. 
BOUSQUET. ' NoHveait iraiU de la vaccine^ p. 403 et seq. — 3. 
Crookshank. History and Pathology of Vaccination^ vol. ii. — 4. 
De Carro. Hisioire de la vaccine, 1804, p. 23 et seq. ; London 
Med. Gazette, vol. xxix. p. 385. — 5. Murphy, Shirley. Report 
of Med. Officer to Local Gov. Board for 1882, p. 35.-6. Ring. 
Treatise on Cow-pox, p. 20. —7. Saccq. Tmttato di vaccina- 
zione, pp. 145-148. — 8. Final Report of the Royal Commission on 
Vaccination, 1 8g6, sections 19-27 and Appendix J., K " Wood- 
ville's cases." 



During the evolution of the local changes which result 
from the insertion of vaccine lymph beneath the surface 
of the skin, it is possible to recognise three more or less 
definite stages of papule, vesicle, and pustule. 

The same statement holds good with reference to 
the eruption of smallpox, whether this be local, i.e. 
due to inoculation of the virus, or general, as the result 
of infection. 

In each instance the appearance of the first or 
papular stage is brought about by inflammatory re- 
action, causing an increase of intercellular fluid together 
with concomitant increase in volume and number of 
epithelial cells of the rete Malpighii more particularly. 
The papule gradually becomes enlarged by a circum- 
ferential extension of the same process, and owing to 
further changes in the cells first affected. Vacuoles 
arise in the central portion of the papule, by the exten- 
sion of which this ultimately becomes a vesicle. 

The vesicle is a multilocular structure, the dissepi- 
ments, by means of which its interior is divided up, being 

74 VACCINATION: chap. 

formed from the thinned and extended remains of the 
original epithelial cells. Owing to the fact that the 
process of vacuolation, for a time, increases more ex- 
tensively at the advancing edge of the vesicle, the 
central portion remains somewhat less elevated, thus 
giving rise to the appearance termed umbilication. 

Stanley Kent, as the result of histological examina- 
tion of a series of vaccine vesicles, which I had removed 
from the calf at gradually increasing intervals of time 
after vaccination, showed that at a quite early stage of 
the process an outflow of leucocytes takes place, towards 
the point of injury. In time each blood-vessel be- 
comes the centre of an aggregation of leucocytes, which 
by the rapid increase in their numbers eventually trans- 
form the originally clear inflammatory exudation into 
a purulent fluid. The vesicle is said now to have 
become converted into a pustule. 

By the thinning and ultimate rupture of its trabeculse, 
the pustule finally becomes unilocular. The turbid 
fluid contained in it now gradually dries up and, 
together with the necrosed remains of epidermal cells, 
takes part in the formation of the crusty which under 
the microscope appears as a homogeneous mass very 
deeply coloured by the ordinary stains. 

Meanwhile a regeneration goes on underneath the 
crust, the new epidermis being formed by an ingrowth 
from the surrounding stratum lucidum. The extent to 
which the cutis vera has been involved determines the 
depth of the resulting scar. 

The minute local changes produced in the skin of 


the calf by vaccination has recently been made the 
subject of careful study by Dr. Gustav Mann, who also 
employed for the purposes of his research specimens 
of skin removed by me at varying intervals after the 
operation of vaccination. His description of the 
normal skin of the calf, and of the changes produced 
in it by vaccination, is based on the examination of 
material which was fixed, immediately after removal, in 
picric acid and corrosive sublimate mixture and cut in 

The abdominal skin of the calf exhibits a sharp 
demarcation into epidermis, dermis, and hypodermis. 

The epidermis is composed to one-fifth of its thick- 
ness of the stratum corneum, then follows a single 
layer of cells with the characteristic granules of the 
stratum granulosum, and finally the stratum Malpighii, 
built up of cells three to four layers deep. Of these 
cells those nearest the dermis have long finger-like 
processes fitting into a very dense layer of white fibrous 
tissue. Weigert-Kromayer's staining method reveals 
fibrils which in the youngest layer of epithelial cells are 
arranged vertically, while in the second and subsequent 
strata they assume a horizontal course, giving rise to 
the "prickly" appearance and uniting the cells very 

The dermis is composed (i) of the already men- 
tioned dense outermost layer [basal membrane] to 
which the epithelial cells are attached; (2) of a very 
loosely arranged connective tissue, consisting of fine 
bundles of collagenous fibrils with clasping cells, which 

76 VACCINATION: chap. 

latter lie with their wing-like processes parallel to the 
surface of the skin ; and (3) of an inner elastic zone 
distinguished by the presence of coarse elastic fibres 
which for the most part run horizontally. These fibres 
give off fine twigs, which pass obliquely upwards into 
the papillary zone, and also form densely felted 
cylinders in the outer root sheath of the hairs. 

The hypodermis is chiefly composed of thick bundles 
of collagenous tissue, with a few elastic elements in the 
septa separating the bundles one from another. In 
the external region of the hypoderm is found a plexus 
of very large lymphatics. 

Vaccination, in all the specimens examined, has 
resulted in a complete division of the epidermis, the 
vaccine lymph employed being therefore brought directly 
into contact with the loose dermal tissue, and it will be 
seen that the chief changes are brought about in that 
zone which lies between the hypoderm and the basal 

In a specimen removed within an hour after vaccina- 
tion, the wound is blocked by a clot which externally is 
of a coarsely granular nature ; between the edges of 
the epidermis it is finely granular, and in the dermis is 
found a close fibrin reticulum, denser at its periphery 
than in the centre, and enclosing numerous red and a 
few white corpuscles. The blood-vessels close to the 
injury are dilated, and many of them completely throm- 
bosed with a mass of white corpuscles. That already 
some change has taken place in the vessel walls is 
evidenced by the fact that close to the site of injury 


red corpuscles may be seen adhering to the lumen of 
the capillaries and arterioles, while the blood from the 
rest of the specimen has escaped at the time of removal 
of the skin. 

The nuclei of both the epidermal and dermal cells 
in the neighbourhood of the lesion are swollen, and the 
basophil chromatin contained within them is increased 
to at least double its normal amount. 

In the dermis an emigration of leucocytes into the 
loose connective tissue spaces is noticeable. It is 
possible to demonstrate at this very early period" a 
considerable number of diplo-bacilli and single very short 
bacilli in that part of the clot which lies between the 
edges of the epidermis and also in the somewhat 
cedematous connective tissue clefts quite close to the 

After twenty-four hours the epithelium close to the 
injury has increased two to threefold in thickness, and 
already a phenomenon, which gradually becomes more 
and more pronounced up to the seventy-second hour, is 
seen, namely, the formation of Guarnieri's supposed 
parasites. In methyl blue eosin- stained sections the 
bluish nuclear chromatin is increased, but even more 
so the nucleolar chromatin, which stains a bright red 
colour with the eosin. By a process, which may be 
studied under normal circumstances in the pancreas, a 
considerable portion of this nucleolar matter leaves the 
nucleus and is found lying free in the cytoplasm ; in 
some cells one only, in others several granules are 
extruded. These are at first very minute, but, later. 

78 VACCINATION : chap. 

they increase in size and occasionally seem to fuse ; 
in this way there arise more or less solid spheres lying 
alongside the nucleus, or even indenting it. 

From the twenty-fourth to the forty-eighth hour an area 
varying in width from one to two millimetres has become 
affected. The dermis shows a gradually increasing 
oedema, accompanied by an emigration of leucocytes, 
which results in a tumefaction of the outer dermis, 
rendering its structure even looser than normal. The 
deeper portions of the dermis, so rich in elastic fibres, 
show as yet practically no changes, and the hypodermal 
cells, except for a somewhat greater affinity for basic 
dyes, are also unaltered. As the result of this 
oedematous condition, developed in the neighbourhood 
of the site of inoculation, the lymph is prevented from 
escaping downwards by the dense elastic layer of the 
dermis and the thick collagenous bundles of the hypo- 
derm. Towards the periphery the lymph channels are 
blocked by leucocytes, and there remains only one 
path open for the lymph, namely, through the basal 
membrane into the intercellular channels between the 
epithelial cells. There results thus a widening of the 
normal lymph channels up to the stratum corneum, 
and if it were not for the very resistant horny layer and 
the dried-up clot at the site of the incision, the lymph 
would readily escape in this direction. 

It may be noted that during the second day many 
leucocytes are seen between the epithelial cells, differing 
from those found normally in being usually more or less 
spherical, rather than angular and elongated. 


The changes most characteristic of vaccination are 
found during the third, fourth, and fifth days (72 to 
120 hours after vaccination). 

At the end of three days one may, for descriptive 
purposes, distinguish several zones. Furthest away 
from the line of inoculation the only change noticeable 
is a dilation of the inter -epithelial lymph channels. 
Immediately within this region all epithelial cells, except 
those of the stratum corneum, are much swollen and 
contain granules giving the same reactions as those 
found in normal cells of the stratum granulosum. We 
find thus a premature ageing of the cells. 

The dermis underneath these cells forms large 
bullae, the walls of which consist of compressed con- 
nective tissue cells, collagenous fibres, and numerous 
leucocytes. No wandering cells are found in the bullae, 
but fairly numerous diplobacilli and single bacilli. 

Still nearer the point of inoculation the epithelial 
cells contain markedly enlarged nuclei, some of which 
before division reach a diameter two to three times 
greater than normal. As a rule, the nucleus fragments, 
however, into six to twelve smaller nuclei, which, lying 
closely together, become flattened against one another on 
one or more sides. Concurrently with the formation of 
these multi-nucleated giant cells one meets also with 
greatly distended lymph vesicles in the epithelium, the 
walls of which are composed of the enormously stretched 
fibrillse of degenerated epithelial cells. In the interior 
of the vesicles, in addition to the fibrin reticulum, are 
found various micro-organisms. 

8o VACCINATION: chap. 

Internal to the zone just described the giant cells 
are replaced by cells having only a fifth to a quarter the 
size of the multi-nucleated cells and possessing only one 
or two nuclei. These smaller cells are spherical, while 
the larger ones are angular in outline. It seems prob- 
able that the smaller globular epithelial cells are 
derived from the giant cells. 

In the centre of the vaccinated area no living 
epithelial cells are present, but only remains of the 
stratum corneum and of a dense dried-up blood-clot. 

The changes described for the third day also hold good 
for the fifth, the only difference being that the central 
necrosed area has increased in size and that the zone 
of infection has spread out laterally. By the continued 
emigration of leucocytes the central zone necroses more 
and more, the connective tissue elements succumbing 
to the pressure exerted by the wandering cells. The 
only elements which do not undergo any change during 
this death of the dermal elements are the elastic fibrils, 
which up to the very end stain quite sharply with orcein. 

While up to the end of the second day practically 
no changes could be made out in the hypoderm, there 
is found later on, and especially about the fifth day, a 
considerable swelling of the thick white fibrous bundles, 
called forth by the great activity of the fixed connective 
tissue cells. These latter stain more deeply with basic 
dyes than normal cells, and their numerous branching 
processes will stand decolourisation more strongly. 

No reference has been made, up till now, to what I 
consider the most characteristic feature of vaccination, 


viz., certain appearances which are constantly seen in 
tissues fixed by picro-corrosive subhmate solution, and 
also, but not so well, when alcohol is employed for 
fixation. On staining sections in Loffler's methylene blue 
(in which the normal amount of caustic potash is 
increased ten times) for twenty-four to forty-eight hours 
at 30° C, and then differentiating for ten to thirty 
minutes in i per cent acetic acid, one finds immediately 
outside the necrosed area, in the superficial loose dermal 
tissue, a number of globular masses, varying considerably 
in size, and arranged either singly or in pairs. 

The further outwards the smaller these globules 
become, till at the spreading zone one finds in the con- 
nective tissue clefts very short bacilli arranged singly or 
in pairs. 

The idea that the larger globules represented either 
a capsulated, sporulated, or involuted stage of the 
bacillus led to the endeavour to demonstrate if possible 
a central corpuscle. After many failures the following 
method has been found efficient for this purpose. 
Sections must be stained in Unna's polychrome methy- 
lene blue for twenty minutes, then be differentiated by 
Unna's method in 30 per cent tannic acid for one to 
three minutes, according to the thickness of the section. 

Although the globules vary in diameter, the stainable 
spot in the interior, which appears spherical or slightly 
elongated, is usually of the same size as the micro- 
organisms in the peripheral advancing area. 

The exact nature of these bodies is being made the 
subject of further investigation. 


Chemistry and Morphology of Vaccine Lymph 

Fresh vaccine lymph, taken before full maturity of 
the vesicle, is a clear, transparent, limpid fluid, almost 
colourless in man, and slightly straw-coloured when 
obtained from the calf; this difference in colour depends 
on the varying quality of the normal pigment present in 
the blood plasma. Calf vaccine lymph is also some- 
what more viscid than human lymph, and does not flow 
so readily when the vesicle is punctured, for which 
reason it is necessary to use compression forceps in the 
ordinary methods of collection of calf lymph. 

Examined microscopically, vaccine lymph contains a 
certain amount of epithelial debris, a few cells and 
portions of cells being always visible. Leucocytes are 
usually present also, the number depending on the 
stage at which the lymph is taken ; few or even none 
are to be found in the contents of the vesicle when first 
formed, but at or after the period of maturation they 
may be so numerous as to render the lymph turbid, or 
even puriform. 

A few red blood corpuscles may be noticeable, 
although most observers will hardly agree with the 
statement made by Dr. Husband to the Royal Commis- 
sion on Vaccination that this is invariably the case. 

In stained specimens particularly, micro-organisms 
of one or more varieties can be readily demonstrated, 
the actual numbers being dependent to a certain extent 
on the care with which the lymph has been collected. 


The nature and significance of the various microbes, 
which at one time or another have been isolated from 
specimens of vaccine lymph, will be discussed in the 
section on bacteriology. 

It is matter of common knowledge that untreated 
lymph, when stored in capillary tubes, tends after a 
longer or shorter interval to become cloudy, under 
which circumstances it is also liable to be uncertain in 
its action if subsequently used for the operation of 

This opaque appearance may be quite independent 
of any coagulation of the lymph, as may not unfre- 
quently be demonstrated on breaking tubes in which 
it is most marked. On the other hand, where clotting 
has taken place after the lymph has been stored, 
the opacity tends to form with the coagulum a central 
whitish thread in the midst of a clear fluid, instead 
of being distributed through the lymph in discrete 
points, as may otherwise be the case. 

If cultivation experiments be carried out by inocula- 
tion on nutrient media, the number of colonies resulting 
from such inoculation with the contents of tubes which 
have become opaque is usually much greater than if 
fresh lymph is employed in a similar manner. We are 
apparently justified, therefore, in considering that the 
opacity of such old stored lymph is, in the main, the 
outcome of an enormous multiplication of aerobic 
bacteria, the ancestors of which are present in the 
lymph when first collected, although their numbers are 
then so comparatively small as not to render it in any 

84 VACCINATION chap, iv 

way turbid. It follows that ordinary vaccine lymph, 
which has become opaque, should never be employed 
for vaccination. 

Vaccination lymph, chemically speaking, consists 
of the serum, or rather of the plasma of the blood. 
When freshly obtained, therefore, it is faintly alkaline 
in reaction, but it tends to become distinctly acid 
after a time when stored. In addition to the various 
salts and proteids normally present in the blood plasma, 
vaccine lymph contains some substance, possibly of 
the nature of a toxin or ptomaine, which results from 
the ^dtal activity of the specific organism peculiar to 
vaccinia. That this is so may be proved either by 
filtration of the lymph through porcelain, or by exposure 
of it to a temperature of about 50° C. If, as I have 
shown, such filtered or sterilised lymph be inoculated on 
the skin in the usual manner, no obvious effect is pro- 
duced at the point of inoculation, but it not unfrequently 
happens that the animal has been rendered temporarily 
immune to the effects of subsequent vaccination with 
lymph of normal potency. (See also Smallpox Antitoxin, 

P- ^zz) 

Histology of Smallpox Vesicle : i. Auspitz and Basch. 

Virchow's Archiv, Bd. 28, p. 337 et seq. Of VaCCine Vosicle : 
2. Keirle. Maryland Medical Journ. Baltimore, 1880, vol. 
vii. p. 169. — 3. Kent, Stanley. British Med. Journ. vol. ii. 
Sept. 22, 1894, p. 633. 



Although the study of the bacteriology of vaccinia 
and variola has, during the past thirty years or so, 
attracted the attention of a great number of observers, 
much yet requires elucidation before we can look upon 
our knowledge in this direction as satisfactory. Never- 
theless progress, though slow, is perhaps sure, and it 
would seem as if it must be the very difficulties of the 
subject which enhance the inducement for the con- 
stantly renewed initiation of investigations by observers 
in all parts of the world. 

As long ago as the year 1809, Sacco placed on 
record the fact that in vaccine lymph there might 
be found certain granules generally collected together 
into masses, and possessed of the power of automatic 
movement. The observation appears to have stood 
alone for more than half a century, but in 1863 
Dr. Lionel Beale gave an impetus to the investi- 
gation of the morphological constituents of vaccine 
lymph by his description of the existence in it of 
transparent or hyaline particles of extreme minuteness. 
Both in his original paper in the Microscopical Journal 


for April 1864, and also subsequently in the Report of 
the Royal Commission on Cattle Plague, Dr. Beale 
expressed the opinion that these granules constituted 
the contagious principle of the disease, but he does not 
appear to have obtained experimentally any facts in 
support of his belief. 

A few years later Hallier and Zurn (1867) and 
Keber (1868) called attention to the presence in 
vaccine lymph of numerous micrococci, or " grains " as 
they were termed by the latter observer, who also evi- 
dently regarded the bodies found by him as the carriers, 
if not the actual generators, of the virulent principle. 

It is, however, to M. Chauveau (1868) and Professor 
Burdon - Sanderson that we owe the first real step 
toward the elucidation of the micro-pathology of vac- 
cinia, these observers having demonstrated that vaccine 
lymph, when freed from its contained particles, and in- 
oculated on a living animal, no longer causes vaccinia ; 
while, on the other hand, the precipitate or deposit, 
separated from it in one way or another, when em- 
ployed in similar fashion, remains capable of producing 
the disease. 

Chauveau found that vaccine lymph, taken at the 
stage of maturation of the vesicles, contained minute 
granular bodies, to which he gave the name of corpuscles 
noirsj and not unfrequently also much larger bodies 
resembling the leucocytes of pus. He further found 
that if the lymph was diluted ten times, and allowed 
to stand, these leucocytes, when present, gradually sub- 
sided to the bottom of the containing vessel, and 


could then be removed. When this was done, and the 
material employed for an experimental vaccination, a 
local result was still obtained, thus showing that the 
active principle was contained not in these leucocytes, 
but either in the granules or in the fluid portion of 
the lymph. 

In order to settle this point, Chauveau devised a 
method of diffusion by which he was eventually able 
to separate the granules from the fluid portion of the 
lymph. The amount of vaccine lymph at his dis- 
posal being necessarily exceedingly small, the ordinary 
means for obtaining diffusion were not applicable. 
Chauveau attained the desired object, however, by 
carefully floating a layer of distilled water on to the 
surface of a drop of lymph contained in a tiny 
glass vessel of cylindrical shape. If this manipula- 
tion is dexterously performed, the two liquids do not 
mix with each other excepting in the immediate neigh- 
bourhood of the point of junction of the two layers. 
But after a time the soluble constituents of the vaccine 
lymph pass upwards into the water, so that if, after 
twenty-four hours or so, a portion of the upper layer be 
carefully removed by means of a capillary tube, and 
examined under the microscope, it is found to be free 
from "granules." And further, Chauveau was enabled 
to demonstrate the fact that while this upper layer of 
fluid, which could be shown to contain albumen, and so 
afforded evidence of diffusion having taken place, gave 
negative results only on inoculation, the lower layer, 
on the contrary, which contained the "granules" in 


suspension, was found to be active as vaccine, normal 
vesicles being obtained as the result of its inoculation. 

Professor Burdon- Sanderson shortly afterwards re- 
peated Chauveau's work, employing certain improvements 
in the method which seemed likely to render it more 
accurate, and thus to afford results of greater scientific 
value. He also made use of another method of his own, 
consisting in the filtration under pressure of the liquid 
to be tested through a layer of unglazed porcelain, by 
which means all solid particles were effectually separated 
from the fluid containing them. Professor Burdon- 
Sanderson, by the employment of this method, and 
also of his modification of that originally introduced 
by Chauveau, fully confirmed the results announced 
by the French investigator. 

Further, Chauveau, by largely diluting vaccine lymph 
with distilled water, and employing the mixture for 
vaccination, found that he thus obtained strong addi- 
tional evidence in support of the doctrine, now univer- 
sally accepted, that contagium is particulate in nature. 
He was enabled to show that whatever the degree of 
dilution, the local effect produced (provided that it is 
produced at all) is always the same. In other words, 
the difference between the effect of inoculating extremely 
diluted lymph, and that of inoculating the lymph itself 
without any addition of water, does not manifest itself, 
as might be expected, in the size of the individual 
vesicle or vesicles produced. He found, on the other 
hand, that the difference consisted entirely in the 
numerical relation of successes to failures. If success- 


ful, inoculation with a liquid containing an extremely 
small proportion of lymph may communicate the disease 
as completely as the original lymph itself, a result which is 
not only perfectly consistent with the fact that contagium 
is particulate, but which is in itself a necessary conse- 
quence of its being so. 

As bearing on this question, it may be mentioned 
that Schultz of Berlin has quite recently been en- 
gaged on an investigation, having for its object the 
determination of the extent to which, in order to save 
time and expense, glycerinated vaccine lymph could be 
diluted without losing its efficiency as vaccine. In the 
course of his experiments he found that an emulsion 
containing one part only of vaccine pulp in two thousand 
parts of glycerine and water, was capable of producing 
locally a normal vesicle in a small proportion of the 
cases in which it was employed for vaccination. 

The experiments of Reiter confirmed those of 
Chauveau, and showed that vaccine lymph, even when 
excessively diluted, was capable of affording positive 
result on vaccination, provided the more extensive the 
dilution the greater the absorbing surface. 

In 1872 a paper was published by Cohn of Breslau, 
in which he treated the morphological aspects of the 
subject with much completeness. His observations, 
which related to both vaccine lymph and variolous 
lymph, have received general corroboration from all 
subsequent workers. He, however, apparently believed 
certain micro-organisms found by him to be of one 
species only, to which accordingly he gave the name 


micrococcus vaccinae or variolas, as the case might be, 
whereas later observers have shown that organisms of 
more than one species are usually to be found in any 
given specimen of crude lymph. Cohn called attention 
to the fact that in perfectly fresh vaccine lymph the 
" corpuscles " for the most part occur singly, but that 
others are joined together in pairs in a form resembling 
the figure 8 ; and he states that after the lymph has 
been kept for a time the number of the " double cells 
increase, and soon chains of four begin to be distinguish- 
able. These chains are usually curved or in zigzags ; 
their attachment one to another is evidently very slight, 
as they can readily be displaced. . . . After a few 
hours' observation they are seen to be all aggregated 
into irregular colonies or clumps, each consisting of 
sixteen, thirty-two, or more corpuscles." He also noted 
a point of importance in connection with the opacity 
which is apt to occur in stored lymph, namely, that "in 
capillary glass tubes the enlargement of colonies some- 
times lasts a long time, so that groups of them acquire 
considerable size, and present themselves as flocculi." 

Klebs (1873) isolated from vaccine lymph micrococci 
united together in the form of tetrads, which he con- 
sidered as specific, and to which he gave the name of 
tetracoccus vaccinae. Cornil and Babes (1883) also 
met with micrococci. Koch (1882) found micrococci in 
the vaccine vesicles of a child, and noted that human 
lymph, freshly prepared with glycerine, contained what 
he appears to have regarded as the specific bacteria of 
vaccine. These developed but slowly, even in the 


incubator, and were met with in fewer numbers in old 
lymph, even though it still remained active. 

Quist in the following year (1883) published a series 
of experiments dealing with the possibility of cultivating, 
outside the animal body, the micro-organisms present in 
vaccine lymph. The culture fluid employed by him was 
composed of equal parts of blood-serum, glycerine, and 
distilled water, this mixture being rendered alkaline by 
the addition of -g-^^th part of carbonate of potash. After 
sterilising this fluid by exposing it to a temperature of 
60° C. for one and a half hours on three successive 
occasions, it was inoculated with a minute piece of 
sterilised sponge soaked in clear lymph, or with a piece 
of vaccine "crust," which had been washed in distilled 
water and then carefully dried. He found that growth 
eventually occurred both on and below the surface, the 
former consisting of minute scales, while the latter 
gradually settled to the bottom of the vessel as a fine 
sediment. The scales, forming a scum on the surface 
of the fluid, he found to be composed of swarms of 
micrococci, which, when inoculated on the skin of 
calves, in some cases gave rise, in his opinion, to a 
typical eruption of vaccinia. Although Quist did not 
attain any great measure of success in the immediate 
object of his inquiry (the identification of the vaccine 
organism), still he showed that the specific contagium 
of vaccinia could exist for a time, at any rate, in a fluid 
containing a considerable portion of glycerine. 

About this same time papers were published by 
Feiler (1883), Serebriakofl" (1884), Baragi (1884), and 


Hubert (1884), all of whom denied that vaccine lymph 
contained any specific microbe whatever. Voigt isolated 
three species of bacteria by making gelatin plate cultures 
of vaccine lymph. Of these one, which alone was 
constantly to be found, he considered to be specific, 
and consequently termed it the vaccinococcus. He 
describes this as a small coccus growing usually in 
groups of two or four, sometimes in masses, and, less 
often, in chains. These bacteria gave rise to glistening 
white circular colonies, which did not liquefy the gela- 
tin in which they were grown. He further stated that 
cultivations of this bacterium, when inoculated on calves, 
rendered them immune to subsequent vaccination. 

Guttman (1886) also found micrococci in the con- 
tents of vaccine vesicles. These were the micrococcus 
pyogenes aureus, micrococcus pyogenes albus, and 
micrococcus viridis flavescens, but none of these is 
specific to vaccinia. 

Marotla (1886) found a coccus (micrococcus tetra- 
gonus) which he regarded as specific. By its use he 
thought that he had succeeded in producing vaccine 
vesicles. From the characters which he assigns to his 
cultivations, it would appear that in reality what he 
isolated was the staphylococcus pyogenes aureus. 

In 1886 Meguin called attention to the fact that 
the number of micrococci contained in vaccine lymph 
increased considerably from the second day onwards 
after the removal of the lymph. 

During the same year (1886) Dr. Buist succeeded in 
isolating from vaccine lymph three varieties of micrococci, 


which, when grown on nutrient media, gave rise to 
colonies of a white, yellow, or orange colour respec- 
tively. All these Buist appears to have considered to be 
essential constituents of vaccine lymph, as is evidenced 
by the fact that he speaks of them as white, yellow, 
and orange vaccine respectively. From specimens of 
variolous lymph he succeeded in obtaining one organism 
only, the colour of which was white when grown on 
solid media. With none of these cultures did he obtain 
any definite result on the inoculation of calves, monkeys, 
or human beings. 

A certain small measure of success appears to have 
attended inoculation experiments in the hands of 
Carmichael (1887), who does not, however, give any 
detailed description of the morphological characteristics 
of the growths which he obtained, and which, by the 
way, although differing one from another in colour, he 
seems to have considered were the product of one 
species of micrococcus only. As was doubtless the 
case with Quist's experiments, in the few instances in 
which true vaccine vesicles resulted at the site of his 
inoculations, success was in all probability due to the 
presence in the material employed of a small proportion 
of the lymph originally used for the seeding of his culture 

Tenholt (1887) appears to have isolated from speci- 
mens of vaccine lymph a dozen different micrococci, 
two bacilli, and two yeasts. Hlava (1887), on the other 
hand, isolated from vaccine vesicles the streptococcus 
pyogenes, the staphylococcus pyogenes, the staphylo- 


COCCUS pyogenes albus, the staphylococcus cereus 
albus, and the micrococcus viridis flavescens. Garre 
(1887) reported the existence in vaccine lymph of two 
bacilli and of a small coccus, which he considered to be 
specific ; this, when inoculated on the calf, produced 
the vaccinal eruption, followed by immunity, while in 
the human being it afforded immunity, although giving 
rise to no eruption. 

Pfeiffer (1887) recorded the existence in vaccine 
lymph of micro-organisms of several different forms, 
including a yeast (named by him sachromyces vaccinae), 
two sarcinge (lutea and aurantiaca), a short bacillus allied 
to proteus vulgaris, but differing from it in not being 
pathogenic, and certain micrococci, of which one, the 
staphylococcus cereus albus (Passet), was of constant 
occurrence, while the staphylococcus pyogenes aureus 
and S. albus were less frequently met with. Pfeiffer, 
however, believes that the specific agent of variola 
and vaccinia is not to be sought for among the 
bacteria, but rather belongs to the class of sporozoa. 
This subject will require further notice later on. In 
addition to the staphylococcus aureus and albus, M. 
Grigorieu (1889) describes a micrococcus vaccinae as 
specific to the disease. This microbe usually develops 
as a diplococcus, rarely forming chains, and grows 
slowly on gelatin, which it liquefies after four to six 
weeks. On the calf it produced a papular eruption, 
which apparently afforded immunity to subsequent 

M. Woitow (1890) succeeded in isolatingfour staphylo- 


cocci — aureus, albus, cereus, and citreus. He affirms 
that the inoculation of a mixture of these microbes 
produced typical vaccinia in the calf. Professor Leoni 
(1890) met with several pyogenic microbes in fresh 
vaccine lymph, notably the staphylococcus pyogenes 
albus, which was invariably present. M. Protopopoff 
(1890) in like manner found it possible to isolate 
several different forms of micrococci. 

As the result of my own work previously to 1891, in 
which year my first paper was published, it would 
appear that there are at least three species of micro- 
organisms, namely, staphylococcus albus, staphylococcus 
pyogenes aureus, and staphylococcus cereus flavus, 
corresponding probably to Buist's white, orange, and 
yellow vaccine, respectively, one or more of which are 
almost universally to be found in every specimen of 
human or calf vaccine lymph examined. Of these the 
staphylococcus albus is usually to be found in the upper 
layers of healthy skin of unvaccinated persons.* In 
addition, I had been able to satisfy myself as to the 
occasional presence of the streptococcus pyogenes. 

I myself have never detected the streptococcus of 
erysipelas, but Dr. Klein has placed on record the fact 
that in one instance he isolated this microbe from a 
specimen of human vaccine lymph, the use of a portion 
of which was believed to have given rise to the occur- 
rence of this disease. Also I have shown that none 
of the microbes ordinarily to be isolated from vaccine 
material can be regarded as specific to vaccinia. 

Professor Crookshank (1891) succeeded in isolating 

96 VACCINATION: chap. 

from various specimens of vaccine lymph, by the method 
of plate cultivation, an immense number of bacteria, 
including micrococci, bacilli, torulae, etc. All of these 
he recognises as well-known saprophytic bacterial forms, 
associated, some of them, with processes of suppuration, 
but none of which, he says, can be regarded as the con- 
tagium of vaccinia, seeing that no single one of them is 
constantly present in vaccine lymph, human or bovine. 

M. Besser (1893) found in the papules of a case of 
smallpox, at the fifth day, a small bacillus, which he is 
disposed to look upon as the specific organism of 
variola. He obtained cultures on various nutrient media, 
coverslip preparations from which always showed the 
organisms arranged in palisade-like rows. 

The late Professor Straus (1893) found that the 
seeding of gelatin plates with fresh vaccine material, 
whether glycerinated or not, gave rise to the appearance 
of numerous colonies of micrococci, among which the 
stapfhylococcus pyogenes aureus and S. albus were most 
frequently met with. When glycerinated lymph which 
had been kept for a considerable period was employed 
in a similar fashion, the number of colonies was greatly 
diminished. The paper, in which he collaborated with 
MM. Chambon and Menard, is illustrated with three 
figures of plate cultures from vaccine lymph, which had 
previously been stored for different periods of time. 

M. Anthony (1893), i" ^^e course of the examination 
of a number of different specimens of vaccine lymph, 
glycerinated and otherwise, found that four different 
kinds of micrococci and three species of bacilli are apt 


to be present. Of these, what he terms, from its appear- 
ance, the "porcelain micrococcus," is invariably present 
in fresh vaccine. The other three micrococci included 
the staphylococcus pyogenes aureus, a grayish -white 
micrococcus liquefying gelatin, and a yellow micrococcus 
often present, which does not liquefy gelatin, and which 
grows easily in peptone beef- broth, on gelatin, and on 
agar. The four bacilli noted by him include the bacillus 
subtilis, the bacillus mesentericus, a fluorescing bacillus, 
and a motile bacillus which grows in the form of yellow 
colonies. All these bacilli, according to this observer, 
represent impurities of the lymph. As the result of his 
researches, M. Anthony concludes that vaccine lymph 
cannot be regarded as above suspicion so long as it is 
found to contain any microbes other than the porcelain 
micrococcus. The inoculation of cultures of his porce- 
lain bacillus on heifers afforded only negative results. 

M. Maljean (1893) published a paper in which he 
describes the occurrence in vaccine lymph of two yellow 
cocci, of which one liquefies gelatin and the other does 
not, and of a peculiar micrococcus which is of a brilliant 
white colour when grown on various media, and to 
which he gives the name of ^''coccus vaccinal.'''' This 
micrococcus is larger than streptococcus pyogenes, and 
in cultures it grows as isolated points, as diplococci, and 
in short chains. Maljean states that this coccus gave 
rise, in the guinea-pig, to the formation of a suppura- 
tion, and on the calf to a typical vaccine eruption. 
Other observers, however, employing the same cultures, 
failed to obtain a positive result. 



Dr. Klein agrees with me in maintaining that none 
of those microbes, which ordinarily can be cultivated 
from vaccine lymph, are "an essential inhabitant in 
vaccine " or *' can have anything to do with its active 
principle." In the Report of the Medical Officer to the 
Local Government Board for 1892-93, he, however, 
described a peculiar, extremely minute bacillus as 
occurring in calf lymph, and in variolous lymph of 
human origin, obtained at an early phase of the dis- 
order; in calf lymph 72 to 96 hours after vaccination, 
in human variola during the third or fourth day. In 
both instances the lymph was collected aseptically. 
Clear lymph only, as free as possible from epidermal 
debris, was used for film specimens, which, after heating 
and treatment with 30 per cent acetic acid for some 
minutes, was subjected to prolonged staining in alcoholic 
gentian violet. Some of the films of calf lymph (collected 
after removal of the epidermis as a whole) showed 
an abundance of these minute bacilli, generally massed 
together, some of the specimens looking indeed like 
film specimens of an artificial culture. Lymph of early 
human variolous vesicles showed similar bacilli, but 
not so abundantly. Calf lymph taken at later stages 
(five or six days old) showed no bacilli or only a few 
here and there. In certain of these bacilli from vaccine 
lymph. Dr. Klein succeeded in demonstrating spore-like 
bodies which, together with the absence of the bacilli in 
the lymph of later stages, led him to the conclusion that 
disappearance of these bacilli from late lymph was 
possibly the result of their having broken up after the 


formation of spores, which might alone be present about 
the time of maturation of the vesicle. Dr. Klein adds 
that these bacilli are incapable of cultivation in the 
ordinary culture media, no growth whatever having been 
obtained by him on soHdified blood-serum, glycerin agar, 
ordinary agar, sugar gelatin, or ordinary gelatin, although 
in specimens of the same lymph as was employed for 
this purpose, and which on transference to the calf pro- 
duced perfect vaccine vesicles, bacilli were found in 

MM. Baillard and Anthony (1894) have worked at 
the question of the number of microbes present in vaccine 
which has been stored for various periods. Baillard con- 
stantly found in the lymph a white staphylococcus, a 
yellow staphylococcus, and bacillus sub tills. His experi- 
ments led him to the conclusion that, by keeping, . the 
number of germs contained in glycerin pulp become 
considerably diminished, but that germs never completely 
disappear. After so long a time as seven months the 
material with which he was experimenting still contained 
some living germs, among which were the bacillus subtilis 
and staphylococcus albus. Leoni (1894), also experi- 
menting on the effect of glycerin in the purification of 
vaccine lymph, found that the microbes usually present 
had disappeared in from one to four months. 

Dr. Buttersack (1894) described the appearance in 
vaccine lymph, dried on cover-glasses, of a dense net- 
work of threads associated with numerous granules, 
which he considered as peculiar to vaccine. These 
threads he regarded as the growing vegetative part of 


a vaccine fungus, the granules in his opinion representing 
spores. It appears fairly certain, however, that what he 
believed to be a mycelium with spores, in reality con- 
sisted merely of threads and granules of fibrin or of 
some other albuminous substance. 

In this same year (1894) I published a description of 
numerous minute bacilli sometimes to be found in con- 
siderable numbers in the vaccine vesicles both of the 
calf and of the human being at an early stage of their 
development, in this respect, apparently, confirming the 
experience of Klein, which was published in the same 
year, and of which mention has already been made. 
Similar bacilH were also demonstrated by Mr. Kent and 
myself in sections of skin passing through the site of a 
vaccine vesicle obtained from the calf. 

An account of certain experiments dealing with the 
possibility of cultivating these micro-organisms, and also 
a similar one isolated from variolous material, I propose 
referring to later. 

M. Ruete (1894) obtained what he believed to be pure 
cultures of the specific agent of vaccine from calf lymph, 
this consisting of a micrococcus, the toxin elaborated by 
which he termed vaccinin. He failed, however, in his 
attempts at inoculation of the human being. 

In 1895 Landmann found that in Germany no less 
than 80 per cent of certain children vaccinated success- 
fully presented an excessive inflammatory condition of 
the arm, sometimes of an erysipelatous or haemorrhagic 
character. In searching for the cause of these unfortun- 
ately numerous compHcations, Landmann undertook an 


elaborate study of the lymphs distributed from thirteen 
of the German institutes. As a result of his research, he 
found that the number of germs present in the various 
specimens of lymph varied from 50 to no less a number 
than 2,500,000 per cubic centimetre. Among the 
microbes which he succeeded in isolating were the 
streptococcus pyogenes, the staphylococcus albus, and 
the staphylococcus aureus. 

Two American observers. Dr. Stephen C. Martin 
and Professor Ernst, recorded in 1895 the isola- 
tion of a micro-organism which they considered to be 
specific to vaccinia. This was a short thin bacillus, 
which they found was capable of growth on serum 
obtained from ox or horse's blood, but not on other 
media. The growth, which is of a white colour, forms 
chains, and, under certain circumstances, according to 
these observers, takes on the form of a micrococcus. 
With a cultivation of this microbe of the fourteenth 
generation they produced a vaccine vesicle on an infant 
in one attempt out of ten ; better success, however, 
attending their attempts at inoculation of a calf. M. 
Le Dantec in the same year (1895) found, in lymph, 
staphylococci, the reactions of which, he says, show 
that they are related to the ordinary pus cocci. These 
staphylococci differed according to the species of animal 
from which they were obtained ; those isolated from calf 
lymph Hquefy serum, while those obtained from human 
lymph did not do so. 

M. Arloing (1896) experimented with the lymph of 
vesicles of horse-pox, which he had obtained by injecting 

y^ CCINA TION : chap. 

a colt intravenously with vaccine lymph purified as far 
as possible by storage in glycerin. From the resulting 
vesicles he first carefully removed the superficial layer of 
epidermis, and then collected the underlying material 
with aseptic precautions. With this he inoculated tubes 
of glycerin beef- broth, obtaining as a result, in each 
instance, a growth consisting of a micrococcus, which, 
however, when inoculated in turn on the animal body, 
produced neither an eruption nor immunity to subse- 
quent vaccination. From his researches Arloing con- 
cluded that the virulent agent of vaccinia must be of 
the nature of a soluble toxin. 

MM. Boureau and Chaumier, also in 1896, found a 
number of different microbes in specimens of vaccine 
lymph, including staphylococcus aureus, cereusj and 
albus, micrococcus flavus, bacillus subtilis, and luteus, bac- 
terium termo, proteus vulgaris, a cladothrix, a fluorescing 
bacillus, and several other bacilli which they were unable 
to identify. They make the extraordinary statement 
that it is only when the vaccine has lost its virulence 
that the microbes originally contained in it completely 
disappear; that in fact the number of staphylococci 
present at any given time afford a measure of the 
potency of the lymph. 

Dr. Saint- Yves Menard (1896), in reply to this com- 
munication, reiterated the contention of Straus and 
himself, confirming my previous work on this subject, 
that the microbes which can be isolated from vaccine 
lymph by the ordinary methods are in no way essential 
to its specific action, and that by the end of six months, 


at any rate, no micrococci can be obtained from 
" glycerinated emulsion," which has been preserved in 
sealed tubes, although it is still perfectly active as 

Dr. Sacquepee (1896), in the course of examina- 
tions of various samples of vaccine tymph, met with 
three species of staphylococcus, the bacillus sub- 
tilis, the bacillus mesentericus, and another unknown 
species of bacillus. Of these he regards the bacillus 
subtilis and the bacillus mesentericus as evidence of 
accidental impurity of the lymph. Delobel and Cozette 
(1896-97) collaborating with Gourny, in common with 
most other observers, found that the bacteria most 
constantly present in vaccine lymph are a white and 
yellow micrococcus. They state that when due 
care is not exercised in the collection of the lymph, 
bacillus subtilis and bacillus mesentericus may be found. 
Inoculation experiments on the human being and on the 
calf afforded them only negative results. 

M. Paul, also in 1896, published a voluminous account 
of his researches on the occurrence and significance 
of pathogenic micro-organisms associated with vaccine 
lymph. He lays stress on his observation that the staphy- 
lococcus pyogenes aureus is by far the most ubiquitous of 
the microbes that are found in vaccine lymph, although 
it is certainly not an essential concomitant. He finds, 
moreover, that it exhibits both variable resistance and 
variable virulence. A sample of lymph, for instance, of 
supposed Jennerian stock he found to contain this 
staphylococcus in a phase of much higher virulence and 


resistance than was a specimen of this same microbe 
obtained from a sample of calf vaccine lymph. This 
staphylococcus, he adds, may be present without causing 
suppuration ; but the use of a lymph containing it in 
large quantities should, naturally, be avoided. Accord- 
ing to this observer, the number of bacteria present in 
fresh lymph appears to be independent both of the 
origin of the stock from which it is derived and of the 
method of vaccination employed. This, as shown else- 
where, does not accord with my experience. 

Quite recently (1897), in -conjunction with Dr. Blaxall, 
I have carried out a further exhaustive series of inves- 
tigations on the bacterial flora of calf vaccine, with the 
outcome that, as the results of former work had led us to 
anticipate, and, as has been repeatedly shown by other 
observers, calf lymph (or rather vesicle pulp) may con- 
tain a large number of micro-organisms which are in no 
way concerned with its specific activity. Human lymph, 
as a rule, we find to contain remarkably few microbes, 
but, probably, this might not be the case were the whole 
substance of the vesicle removed as is now usual in 
collecting from the calf. We have shown also, as have 
other investigators, that lymph, if carelessly collected, 
is often found to be contaminated with many of the 
numerous saprophytes common to dust, etc., in addition 
to certain possibly pathogenic microbes which may be 
present. In the particular series of experiments here 
referred to, and in a very large number of examina- 
tions of samples of calf lymiph, we have met with the 
following micro-organisms at one or another time. 


They are arranged in the order of their prevalence and 

r Staphylococcus cereus flavus^ 
^' I „ „ albus j ^ 



Large yeast, orange coloured. 
Small ,, light-brown colour. 
Small ,, pale salmon colour, and very slow 

3. Staphylococcus pyogenes albus (Rosenbach.) 

4. ,, „ aureus (Rosenbach.) 

5. ,, ,, citreus (Passet.) 

6. Bacillus mesentericus vulgatus. 

7. ,, subtilis. 

8. Moulds, _pem'a7/m, mucors^aspergilli : Sd.rcmsc, /z^fea, 


In the series of experiments here dealt with, one or 
more members of groups i and 2 were always present. 
The slow-growing yeast was not, however, generally 
visible as a distinct colony till the seventh or eighth day. 

Staphylococcus pyogenes albus was frequently present 
in lymph, but generally in small numbers. Staphylo- 
coccus aureus was rather less frequently observed, and 
staphylococcus citreus was only occasionally met with. 

Bacillus mesentericus may be regarded as an 
accidental contamination. 

Bacillus subtilis rarely occurred if precautions were 
taken ; its presence seeming to depend entirely on the 
care exercised in cleansing the skin, and in collecting 
the lymph material. 

The same is true of group 8. 


It must be stated, however, that all the samples of calf 
vaccine examined were derived from the same station, 
namely, the Government Animal Vaccine Establishment, 
Lamb's Conduit Street. Here, calf-to-calf vaccination 
is continuously practised ; so that the adventitious 
saprophytes found in the lymph, at one and another 
time, are apt to be similar in kind, the extraneous 
organisms originally present having been, so to speak, 
cultivated /^n/^^jw with the vaccine virus itself. Very 
probably, therefore, a strain of lymph from another 
station might present a " flora " differing in kind and in 
amount, as, indeed, has been found to be the case with 
certain foreign lymphs which we have had the opportunity 
of examining. 

On the other hand, lymph taken directly from calves 
vaccinated with vaccine material rendered free from 
extraneous organisms, as for instance glycerinated lymph 
a month after emulsification, often presents remarkably 
few colonies of " extraneous " organisms. In one such 
sample only four colonies appeared on a plate-culture of 
the lymph ; one of staphylococcus cereus albus, and 
three of a light-brown yeast. 

If a vaccine vesicle of a healthy child or calf be care- 
fully opened at an early stage with all aseptic precautions, 
and a droplet of the lymph which exudes from it be 
submitted to bacteriological examination, it may not 
unfrequently be found, as Klein has also demonstrated, 
that such lymph is absolutely pure in the sense that it 
contains no micro-organisms which will grow on ordinary 
culture media. Yet it is perfectly efficient as vaccine. 


But, on the other hand, as just stated, in lymph obtained 
in ordinary fashion from mature vesicles, inoculations of 
plates or tubes of nutrient material usually result in 
abundant growths of micro-organisms. From the list of 
species which I have set out, it will be seen that those 
which are most commonly met with are, morphologically 
and culturally, identical with the staphylococci found in 
pus, viz., the staphylococcus aureus, albus, cereus albus, 
and cereus flavus. Of these the first two liquefy gelatin, 
the other two do not. Occasionally the streptococcus of 
pus may be present, as also the staphylococcus citreus. 
Other microbes of a purely saprophytic nature that are 
met with, include the common hay bacillus, bacillus 
mesentericus, varieties of proteus, yeasts, moulds, and 
sarcinae. These, however, can for the most part be 
excluded by careful manipulation, so that in vesicle 
pulp, which has been collected with all due precautions, 
nothing is found beyond some one or more of the four 
first-named staphylococci, of which a white staphylo- 
coccus is perhaps most common, and possibly one or 
more yeasts. Thus our most recent experience confirms 
in all essential particulars my first report published in 

The fact that in vaccine lymph or in the vesicle pulp 
micro-organisms are found which, under certain cir- 
cumstances, are known to be provocative of suppuration, 
does not in any way imply that the lymph in which they 
are present is purulent, or that, if such staphylococcus 
infected lymph were employed for vaccination in the 
human being, suppuration would necessarily ensue. The 

io8 VACCINATION: chap. 

recent Berlin Commission, indeed, report that of eighteen 
samples of lymph examined only five contained staphylo- 
cocci which were pathogenic for small animals, and, 
further, that no harm resulted on the vaccination of 
children with these particular specimens of lymph. From 
this circumstance it is evident that the virulence of 
these micro-organisms for rodents, for instance, cannot 
be accepted as a measure of their power to affect 
injuriously the body tissues in the healthy human 
subject. The streptococcus of pus must be a rare 
inhabitant of calf lymph, the Berlin Commissioners 
indeed stating that they never once met with it in the 
course of the examination of sixty-four samples of 
lymph. In human lymph its occurrence is by no means 
so unfrequent. 

With reference to the question of the possible pres- 
ence of the tubercle bacillus in lymph, I am not aware 
of any authentic record of its having been found, even in 
lymph derived from the calf. But, as I shall presently 
have occasion to show, we are enabled by the use of a 
special method of preparing and testing vaccine lymph 
to obviate any possibility of danger to which the presence 
of the tubercle bacillus or other pathogenic microbes 
in the original specimen of lymph might conceivably 
give rise. 

In 1894, as already mentioned, I showed concurrently 
with Dr. Klein, though independently, that in specially- 
stained specimens of vaccine lymph taken at a period 
antecedent to full maturity of the vesicles, the presence 
of bacilli of extremely small size and in practically pure 


culture can be demonstrated. These bacilli cannot be 
found, or only with difficulty, in mature lymph, for the 
reason probably that they have by then given place to 
spores. For some time it appeared impossible to obtain 
further evidence as to the role of these bacilli, as 
they altogether refused to grow on any of the ordinary 
culture media, and under either aerobic or anaerobic 
conditions, though this very fact obviously tends to 
show that they are not of a merely saprophytic nature. 

In the following year, however, I carried out certain 
experiments which appear to prove that it is possible to 
obtain a growth of similar bacilli from variolous material 
in a particular culture medium, and even to carry on 
such growth in secondary cultivation. The experiments 
which have been instituted in this direction are but few 
in number, and it is evident that the exact conditions 
essential to success have yet to be determined. 

As it seemed to me, the desideratum for the purpose 
of inducing growth of the supposititious micro-organisms 
was a medium which had not been artificially sterilised, 
which a priori should afford the necessary pabulum, 
and which at the same time was secure in its circum- 
stances against contamination. Hence, the choice 
appeared to He between the intraperitoneal fluids of one 
or another living animal and the hen's egg, which has 
already been made use of as an alternative culture 
medium. As regards intraperitoneal fluid I foresaw 
difficulty, in that it is apt to contain leucocytes, which 
might easily vitiate an experiment, and accordingly 
I adopted the hen's egg as matter of first instance. I 

no VA CCINA TION : chap. 

had no experience to guide me as to the best method of 
carrying out the inoculation of the egg, or as to the 
period of incubation best suited for the object I had in 
view, and my attempts for a time, therefore, met with 
but small success. 

My inoculations were carried out by first cleansing 
the surface of the egg with corrosive sublimate solution 
and afterwards with alcohol, and then breaking a small 
hole in the shell by means of a sterilised needle and 
forceps. Through the opening thus made I mixed the 
yolk and albumen of the egg by means of a platinum 
needle, and then carried out the inoculation by means 
of a glass capillary pipette, which had recently been 
drawn out in the blow-pipe flame. 

For the purpose of such inoculations I employed 
variolous crusts, obtained from the smallpox ships of 
the Metropolitan Asylums Board. These crusts were 
rubbed up in a small glass mortar with a minimal 
quantity of water or sterilised normal saline solution. 
Of the resulting emulsion a drop or two was then taken 
up into the glass pipette and the inoculation carried 
out. Finally, the small hole in the egg was closed up, 
either by means of a small plegget of sterilised cotton- 
wool soaked in collodion or with melted sealing-wax. 

The eggs inoculated in this manner were then placed 
on beds of cotton-wool, and kept in the incubator at a 
temperature of 37° C. Here they were left for varying 
periods of time, a month being the period which was 
eventually found to be that which was most desirable. 

On opening an egg thus treated, I found that in its 


interior was contained a creamy material, which in great 
part had replaced the ordinary egg contents. This 
material, examined in cover -glass specimens, appeared 
to contain a pure culture of one organism only, namely, 
a bacillus which morphologically was not to be dis- 
tinguished from the bacillus which I have previously 
described as to be found in early vaccine lymph. 

My next step was to attempt to grow it in sub- 
culture, and to make observation also of the effect of its 
direct transference by cutaneous inoculation into the 
calf. As regards attempts at sub-culture in all media 
most usually employed for such purpose, including 
gelatin, sugar -gelatin, agar, glycerin -agar, sugar-agar, 
and serum, these at this time failed, as sub-culture of 
the bacilli contained in vaccine lymph had always 
failed when the ordinary culture media are adopted. 
But with the calf it was different, and, as the result of 
inoculation of egg-cultures on this animal, I obtained a 
strain of lymph which, after being passed through a 
series of calves, was successfully employed for the 
vaccination of children. The calves were all subse- 
quently vaccinated with negative results. 

The method of experimentation was as follows : — 
Experiment i. — On 26th March 1895, calf A was 
inoculated by eighteen incisions (fifteen on the abdomen 
and three on the scrotum), with a small quantity of the 
creamy emulsion contained in ^g<g No. 5, which had 
been incubated for one month after inoculation with 
smallpox crusts, as already described. On 30th 
March, scrapings were taken from the vesicular points 

VA CCINA TION : chap. 

which had appeared in the line of two of the abdominal 
incisions, and also from one of the incisions on the 
scrotum, which had become slightly raised and red. 
With material thus obtained, calf B was on the same 
day inoculated by six linear incisions on a specially 
shaved area on the abdomen. Three incisions were 
inoculated with scrapings taken from the vesicular 
points, and three with scrapings taken from the 
incision on the scrotum of calf A. Three days later 
(on 2nd April) all six insertions appeared to be taking, 
though eventually one died away. On 4th April the 
calf was photographed, and on the same day a child 
was vaccinated from it by Dr. Cory. This child I 
have since seen. It has five good marks, and the 
mother told me that the vaccination "did splendidly," 
and that she had had no trouble with the child. From 
calf G, calf H was inoculated on 25th May; and, 
further, this strain of variola -vaccine, derived in the 
first instance from the egg culture, was carried on 
through two additional removes in the calf, after which 
it was not further cultivated. 

Eight calves in all were inoculated in this manner, 
and on 28th May 1895, lymph taken from vesicles at 
the third remove from the egg culture was, in my 
absence, employed for the vaccination of a large 
number of children at the Animal Vaccine Establish- 
ment, owing to the fact that for some reason or other 
the current strain of lymph had failed to give good 
results in the two calves which had been vaccinated 
according to the usual routine. All these children 


showed excellent marks when inspected a week later, 
and several of them were photographed by my colleague, 
Mr. Kent. 

Interesting, and indeed satisfactory, as are these 
experiments from my point of view, I do not claim for 
them demonstration of the practicability of growing 
and multiplying outside the animal body, in artificial 
media, the particulate cause of smallpox, and of con- 
verting the latter by my processes into the essential 
cause of vaccinia. And for this reason : these experi- 
ments of mine on the bovine animal had to be con- 
ducted — at any rate, in this preliminary stage — at the 
Animal Vaccine Establishment in Lamb's Conduit 
Street, and the subjects of them had to be calves that 
on each occasion of experiment by me were at the 
same time vaccinated in some part of their bodies with 
the calf lymph current at this station. It is true that 
every precaution was taken in the way of sterilising 
instruments and cleansing operation tables ; in select- 
ing a given and clearly defined area on each animal 
for insertion of my lymph ; and for keeping the incisions 
made by me protected, as far as possible, against later 
accidental mediate inoculation with lymph from some 
part of the same or other calf into which Animal 
Vaccine Establishment material had been inserted. 
Nevertheless, it is possible that, in spite of all pre- 
cautions, accidental inoculation of my incisions with 
Animal Vaccine Establishment lymph "may have oc- 
curred ; and, further, I am not able to deny that the 
fact of simultaneous inoculation of the calves with my 


114 VACCINATION : chap. 

egg culture and with ordinary vaccine lymph may not 
have, in some occult way, influenced the manifestation 
witnessed on the areas wherein my egg lymph was 
inserted. So, of course, these experiments have to be 
repeated and are, indeed, in course of repetition. 

More recently, Dr. Blaxall and I have been further 
investigating the possibility of the cultivation in artificial 
media of the micro-organism specific to smallpox and 
vaccinia respectively. Our work received considerable 
impetus by the discovery that certain culture -tubes, 
which had been inoculated with vaccine lymph, but 
which appeared to remain completely sterile, neverthe- 
less contained a growth of a microbe indistinguishable 
morphologically from that which I found in egg 
cultures. This growth, although invisible to the naked 
eye, became obvious when cover-glass impressions were 
taken from the surface of the nutrient medium and 
afterwards stained and examined with the microscope. 
Dr. Blaxall subsequently devised a method by means of 
which pure cultivations of apparently the same organism 
were obtained on the surface of agar plates, and this 
not only when vaccine lymph, both of human and 
bovine origin, was employed, but from variolous lymph 
as well. From these plates agar tubes and tubes of 
peptone water were also in turn inoculated. In these 
again, in the first removes, practically no growth was 
visible, except on careful examination by reflected 
light ; but as generations further removed from the 
original stock were obtained, the growth became more 
obvious, showing as a haziness in the fluid media, and 


as a semi-translucent grayish-white hne on a sloping 
surface of agar. Broth cultures of the third and fourth 
generations, when employed for the inoculation of a 
calf, afforded results which we regarded as most en- 
couraging, seeing that we were enabled to carry on the 
infection to other calves and subsequently to children, 
in all of which it took the form of typical vaccinia. 
Some weeks after, the calves were vaccinated with 
active calf lymph, but without result. A later series of 
experiments, however, was not attended with similar 
success. The reasons for so great a divergence in our 
results at one and another time are by no means 
obvious, unless possibly the fact of increased vegetative 
function shown in the later sub-cultures of the microbe 
may have had as a concomitant a lessened degree of 
pathogenicity. On this point we are, at present, unable 
to make any definite statement, but, needless to say, the 
investigation is being continued. 

Another direction in which our research work has 
been extended was suggested to us by certain state- 
ments in the "Report of the Berlin Scientific Com- 
mission." In that report an account is given of 
certain experimental inoculations performed by Freyer 
with the juice of the spleen, liver, inguinal and mesen- 
teric glands, and marrow of previously vaccinated 
calves. Using such material, vesicles were produced 
by the employment of almost every organ, which shows 
that, in Freyer's opinion, the vaccine germ must be 
conveyed to them by the circulating blood. This 
general distribution of that micro-organism takes place. 


he states, up to three or four weeks after vaccination, 
but no mention is made by him as to the earhest 
period at which it was found to occur. The greatest 
amount of success was obtained by the use of inguinal 
gland and spleen juice. He further stated that since 
calves were successfully vaccinated with this material, 
the postulate of an absolutely bacteria-free efficacious 
lymph was complied with. 

It is, of course, obvious that where generalised vac- 
cinia occurs as the result of a local vaccination as, 
according to Chauveau, is always the case with solipeds, 
the micro-organism concerned must have been conveyed 
to a distance from the original point of insertion by 
means of the circulation. Presumably this may also 
take place, although perhaps to a more limited extent, 
in that form of the disease in which, as in the case of 
the vaccination of the human being or the calf, the 
resulting disorder remains to all appearance strictly 
localised, a secondary eruption being prevented by the 
spleen or other organ, such as the liver, being enabled 
to seize upon and ultimately destroy any of the specific 
germs that happen to be carried thither. 

It seemed to me that if Freyer's results were capable 
of confirmation, the possibility of vaccinating by means 
of spleen -juice, for instance, must depend on the 
employment of such material at a period sufficiently 
late to ensure the active principle having reached the 
particular organ, and antecedent to that at which it 
would have been destroyed. I thought also that, if 
the most favourable period could be determined, it 


would be possible then to obtain a material free from 
bacterial growth other than that on which its potency 
for vaccination depended — a material, therefore, which 
would afford exceptionally favourable opportunity for 
the isolation of the organism specific to vaccinia. But, 
unfortunately, notwithstanding the categorical state- 
ments in the Report of the Berlin Commission, we 
have been unable, as the outcome of some preUminary 
experiments, to obtain results comparable to those 
described by Freyer. Neither have we been able to 
demonstrate the presence of a specific micro-organism 
in culture media inoculated from the fresh spleen, liver, 
or glands, obtained at various dates after vaccination, 
or in the interior of fresh or hardened sections of these 
same organs. 


Owing no doubt to the fact that hitherto it has 
appeared practically impossible to isolate from the lymph 
of vaccinia or variola any bacterium which could with 
reason be regarded as specific to these affections, a 
number of observers have turned their attention to the 
search for a micro-organism of somewhat higher type, 
which might by chance be concerned in the dissemina- 
tion of vaccinia and variola respectively. Among the 
first experimenters in this direction must be mentioned 
Renault of Lyons, in 1881 ; Van der Loeff in 1886, 
and Hlava and Pfeiffer in 1887. It was not, however, 
until the publication in 1892 of Guarnieri's work that 
interest in the problem was seriously aroused. Since 


then the literature of the subject has expanded enor- 
mously, owing mainly to the labours of numerous 
Continental pathologists, the only English contributions 
to the mass of experimental work published up to the 
present, being those of Drs. Ruffer and Plimmer and of 
Mr. Jackson Clarke. 

Renault (1881) first called attention to the fact that 
peculiar bodies, which he regarded as parasitic, could 
be demonstrated in the epithelial cells of the vesicles in 
vaccinia and variola. Van der Loeff, also, on examining 
in the warm chamber a hanging drop of clear calf 
vaccine lymph, discovered "numerous small bodies 
capable of independent movement," of which, however, 
he gives no further description ; although he offered the 
opinion that they were living organisms which should 
be classified with the Rhizopoda. 

In 1887 L. Pfeiffer described the occurrence in 
lymph, obtained from cases of vaccinia and variola, of 
small, unicellular, generally spherical bodies, which he 
believed to constitute the specific contagium of these 
diseases. These parasites he found in still greater 
numbers in the Malpighian layer of the skin, multipli- 
cation taking place by division and also by endogenous 
formation of spores. These statements were in part 
confirmed immediately afterwards by Rieck. 

Guarnieri having invariably found, in the epithelial 
cells of the skin in the vesicular stage of smallpox, small 
particles half the size of the cell-nucleus or even smaller, 
next attempted (in 1892) the cultivation of similar bodies 
in the living animal. For this purpose he inoculated the 


cornea of rabbits and guinea-pigs with vaccine lymph, 
with the result that these protozoa, as he considered 
them, appeared in the cells of the epithelium covering 
the cornea. After an interval of about fifty hours from 
the time of inoculation, the surface of the cornea was 
scraped, and the material examined in a hanging drop 
of aqueous humour. Guarnieri also employed for 
examination sections taken through the site of inocu- 
lation and stained in various ways. In either case he 
found numerous " parasites " within the epithelial cells 
of the affected area, generally occupying a zone of the 
protoplasm immediately surrounding the nucleus but 
not intruding upon it. In the hanging drop they 
appeared as Uttle refractile bodies endowed with slight 
amoeboid power. Examination of sections further 
showed that they vary much in shape and size, and, 
further, that they are most numerous in the deep layers 
of the epithelium, the superficial cells not being involved. 
To this "parasite," Guarnieri gave the name of 
Cytoryctes vaccinae. Monti obtained similar results, 
and both Guarnieri and Monti exhibited, at the Inter- 
national Congress of Medicine at Rome, preparations 
in which they demonstrated the presence of the parasite. 
Messrs. Ruffer and Plimmer, who in this country had 
been working at the subject, also attended the Congress, 
and were thus able to compare their own specimens 
with those shown by Guarnieri and Monti, thereby 
establishing, according to Dr. Ruffer, the fact that the 
protozoa described by all of them were identical in 

120 VACCINATION : chap. 

Messrs. Ruffer and Plimmer, writing in 1894, de- 
scribed the alleged parasite as a small, round body, 
which, when treated with appropriate reagents, ap- 
pears to have a more darkly staining centre. It is, 
they say, about four times the size of an ordinary 
staphylococcus, and generally lies in a clear vacuole in 
the protoplasm of the epithelial cells of the stratum 
Malpighii, and occasionally indents the nucleus, although 
they have not found it enclosed in the latter body. 
These observers state that they have found the same 
organism in the sections of skin from smallpox patients 
and in smallpox pustules of the larynx and the trachea. 

In the same year (1894), I.. Pfeiffer not only con- 
firmed the results obtained by Guarnieri, but announced 
also the discovery in the blood of persons suffering from 
smallpox and in the blood of children and calves that 
had been vaccinated, of amoeboid cells which he 
regarded as parasites. These he found to be most 
numerous during the pyrexial stage of the disorder, 
whether variola or vaccinia. These bodies, which are 
somewhat larger in the blood of the calf than in that of 
the child, being in the latter about a quarter the size 
of an average blood corpuscle, are, Pfeiffer states, pro- 
vided with pseudopodia and are occasionally flagellated. 
According to this observer they also contain from one 
to four nuclei, which are readily stained. They occur 
free in the blood, not being contained in cells as is the 
case in the vaccinated cornea ; nevertheless, Pfeiffer 
considers that they represent a similar stage in the life 
history of the organism, of which the more mature form 


is of a cystic nature, to which he gives the name 
Monocystis epithehalis. This cyst he beUeves to give 
rise, in due course, to a number of spores. It should 
be mentioned that he describes similar bodies as occur- 
ring in various other vesicular eruptions of man and the 
lower animals. 

A Japanese observer, Ogata (1895), as the result of 
careful microscopic examination of stained film-pre- 
parations of vaccine lymph, derived from the human 
being and the calf, and also of human variolous lymph, 
concludes that certain protozoa, which he has been able 
to demonstrate in each case, should be classified with 
the Gregarinidae. 

Further confirmation of Guarnieri's experiments was 
afforded in 1895 t>y Von Sicherer, Ernst Pfeiffer, and 
Mr. Jackson Clarke. According to E. Pfeiffer the 
bodies vary considerably both in size and shape, being 
sometimes as large as a red blood -corpuscle. They 
resemble the corpuscles also in their affinity for certain 
staining reagents. 

There can, of course, be no doubt as to the occurrence 
of most of the appearances described, although we may 
not be prepared to accept the interpretation put upon 
them ; and in this connection it is well to bear in mind 
not only that the bodies enclosed in cells, described by 
Guarnieri, Monti, and Ruffer, are said by them to differ 
essentially in their staining reaction and in their appear- 
ance from those of the Pfeiffers, Van der Loeff, and 
others, but also that all the more recent experimental 
work in this direction has been carried out on the cornea 


of the rabbit and the guinea-pig, both of them animals 
which there is reason to believe are insusceptible to 
vaccinia. It is therefore, I think, conceivable that the 
" parasites " in question may represent merely the result 
of epithelial irritation caused by the scarification, 
together with that — of a non-specific nature, however, 
— set up by the vaccine lymph employed. 

That such is indeed the case there is now a consider- 
able amount of evidence to show. Thus Ferroni and 
Massari (1893) expressed the belief that Guarnieri's so- 
called parasites are in reality derived from the nuclei of the 
cells, or are due to the immigration of leucocytes. They 
base their opinion on the results of certain experiments 
devised by them, in which they inoculated the cornea of 
rabbits and guinea-pigs with croton oil and India ink. 
On subsequent examination of stained sectiorLS, they were 
able to demonstrate the presence in the epithelium cells 
of numbers of small bodies apparently identical with 
those described by Guarnieri. It should, however, be 
stated that E. Pfeiffer failed to find them under similar 
circumstances, although they were invariably present 
after the inoculation with vaccine lymph. 

In the An?ials of the Pasteur Institute for April 1897 
appeared a most important paper by M. Paul Salmon. 
This investigator brings to bear on the subject a thorough 
knowledge of modern staining reactions, by the light of 
which he shows that Guarnieri's parasites, while differing in 
their affinity for certain stains from either the protoplasm, 
nucleus, or nucleolus of the epithelial cell, correspond 
in all respects, as judged by such tests, to the nuclei of 


the migratory leucocytes, the chromatin masses of which 
rapidly break down in the substance of the invaded 
cells. He sums up the matter in the following words : 
" Le pseudo-parasite n'est pas une formation endogbne ; 
il a done forcement une origine extra-cellulaire, extra- 
epitheliale, et /a petite masse de chrojnatine ne pent avoir 
qu'une origine; les cellules migratrices.^^ The paper is 
illustrated with drawings showing all stages of the 
process of invasion of the corneal epithelium by the 
leucocytes, from that of their first appearance in the 
underlying tissue, to that of their final dissolution, repre- 
sented by the presence of minute granules of chromatin 
only, in the protoplasm of the epithelial cells. No one, I 
think, comparing this plate with the drawings and descrip- 
tions of the corneal " parasites " of Guarnieri and others, 
can fail to be struck with the truth and completeness of 
Salmon's explanation of the facts previously observed. 

Recent researches have tended to confirm the state- 
ment of L. Pfeiffer as to the presence also in the blood of 
variola and vaccinia, especially during the pyrexial stage, 
of minute, free, granular amoeboid cells. It is, however, 
doubtful whether it is possible to demonstrate a nucleus. 
These cells, as already mentioned, are regarded by 
Pfeiffer as the causative agent of these diseases. 

This question has been investigated in America by 
Dr. Walter Reed (1897), whose attention was directed in 
the first instance to the microscopical examination of 
fresh preparations of the blood of vaccinated monkeys 
and calves, and subsequently to that of vaccinated 
children and smallpox patients. Of eleven vaccinated 

124 VACCINATION : chap. 

animals (nine monkeys and two calves), whose blood 
was examined, Reed found in the blood of seven (six 
monkeys and one calf) "granular amoeboid bodies, 
having a diameter about one-third that of a red blood 
cell " during the active stage of vaccinia. These bodies on 
the decline of the local inflammation again disappeared. 
As showing, however, that these bodies have no causative 
relationship to vaccinia is the fact that Reed occasion- 
ally met with quite similar bodies in the normal blood 
of healthy children and of monkeys. This same observer 
states that pale amoeboid bodies, containing a few dark 
pigment-like granules, may also be present in the blood 
from cases of variola in the human being and the monkey. 
The blood of vaccinated children and monkeys was 
found occasionally to contain bodies of like appearance. 

Reed does not appear to have known of a paper 
published in the previous year (1896) by M. F. Miiller, 
an assistant in Nothnagel's clinic in Vienna. Therein 
Miiller gives a description of "small, generally round, 
colourless granules," which he found constantly present 
in the freshly-drawn blood, both of healthy persons and 
of those suffering from various diseases. These granules, 
which may, he says, be readily distinguished from blood- 
plates, would appear to be of like origin with the minute 
granular amoeboid bodies originally discovered by Pfeiffer. 

Further light has been thrown on this subject by the 
labours of Stokes and Wegefarth (1897), who carried out 
a lengthy series of observations in the bacteriological 
laboratory of the Baltimore Health Department. The 
conclusions at which they ultimately arrived may be 


briefly set out as follows: In the normal blood plasma and 
serum of man and the lower animals varying numbers of 
granules, apparently derived from neutrophil and eosino- 
phil leucocytes, are usually to be found. There is reason 
to beUeve that these granules play some part in bringing 
about agglutination and cessation of motility of many 
pathogenic bacteria and of ultimately destroying them, 
a property possessed by the normal blood. That this 
is so, is judged from the fact that on filtration of dog's 
or rabbit's serum through a Miincke porcelain filter, 
this effect on bacteria was no longer exhibited, although 
the potency of the serum in this direction could be 
restored by adding to it a sediment containing granules. 
Stokes and Wegefarth make also a suggestion which 
may not impossibly account for the undoubted fact 
that, as stated both by Pfeiff'er and Reed, the minute 
granular cells described by them are most readily found 
in the blood during the progress of certain infective 
diseases, although, as Reed shows, they must also be 
regarded as normal constituents of the blood. The 
suggestion referred to is that the bactericidal power of 
both leucocytes and serum in man and many animals is 
due to the presence in them of specific granules, derived 
in either case from leucocytes, which in presence of 
invading bacteria are given up to surrounding fluids or 
tissues. As the outcome of the most recent investiga- 
tions dealing with the question under discussion, it 
must, I think, be admitted that although Guarnieri and 
others who have described the presence of protozoa in 
the blood and tissues of man and the lower animals. 


during certain stages of invasion by variola or vaccinia, 
are undoubtedly correct in their statements as to the 
occurrence of the particular bodies described by them, 
they are altogether mistaken as to the true nature and 
affinities of these bodies. There would indeed appear 
to be good reason for the belief that these so-called 
parasites are in fact nothing more nor less than the 
fragments of amoeboid leucocytes which had originally 
invaded the peripheral circulation and, in turn, the 
particular tissue affected, under the influence of 


Bacteriology: i. Antony. Archives de viedecine militaire, 
1891, 1893, and 1894. — 2. Arloing. "Communication to 
Academic des Sciences," Lyon medical, Mars, 1896. — 3. Boureau 
and Chaumier. Congres de midecine de Nancy, 1896. — 4. 
BuiST. Vaccinia and Variola. 1886. — 5. Carmichael. 
Trans. Philosoph. Sac. of Glasgoiu, 1887. — 6. Chauveau. 
Comptes- rendu s, Ixvi. 1868. — "]. Cohn. Virchozu's Archiv^ 
1872. — 8. CoPEMAN. Trans. Internat. Congress of Hygiene, 
1 89 1, vol. ii. ; Proceedings of Royal Society, 1893; Journal of 
Pathology, May 1894 ; British Med. Jourjt. Sept. 22, 1894, 
Jan. 7, and May 23, 1896. — 9. Crookshank. Trans. Internat. 
Congress of Hygiene, 1891, vol. ii. — 10. Gar RE. Deutsche med. 
IVoch. 1887. — II. GoDLEE. Trans. Pathological Society, London, 
1876-77, vol. xxviii. p. 397. — 12. Grigoriew. Baumgartens 
Jahresbericht, 1889. — 13. Hallier and ZuRM. Virchow's 
Archiv, 1867, — 14. Keber. Virchow's Archiv, January 1868. 
— 15. Klein. Report of Medical Officer to the Local Govern- 
ment Board for 1892-93; (1894.) — 16. Landmann. Hygiene 
Rundschrift, 1895. — ^1- Le Dan tec. Congres de med. interne 
de Bordeaux, 1895. — 18. Leoni. Revista de Hygiene, 1890 and 
1894. — 19. Maljean. Gazette hebdomadaire, 1893. — 20. Paul. 
Oesterreiche Sanitdtswesen, xliii. 22. — 21. Pfeiffer. Zeitschrift 
fiir Hygiene, 1888.— 22. QuiST. Berlin, klin. Woch. No. 52; 
1883. — 23. Sanderson, Burdon. Intimate Pathology of Con- 
tagion ; Thirteenth Report of the Medical Officer to the Privy 


Council. — 24. VoiGT. Deutsche med. Woch. 1885. — 25. 
WoiTOW, Baiimgartens Jahresbericht, 1890. 

Protozoa: 1. Clarke, Jackson. Med. Press and Circular, 
July 25, 1894. — 2. Ferroni and Massari. La riforma 
inedica, 1893, vol. ii. p. 602. — 3. GuARNiERi. Centralb. f. 
Bakt. August 25, 1894. — 4. Ogata. Mittheiluttgen aus der 
fuedicinischen Facultdt der kaiserlich -japanischen Universitdt , 
1895, vol. iii. No. 2. — 5. Pfeiffer, E. Centralblatt f. Bakt. 
und Parasitenkunde, 1895, vol. xviii. p. 769. — j6. Pfeiffer, L. 
Die Protozoen als Kraitkheitserreger, Jena, 1894. — 7- Reed. 
Journ. of Experimental Medicine, Sept. 1897, vol. ii. p. 515. — 
8. RuFFER. Brit. Med. Journal, June 30, 1894. — 9. Salmon. 
Annates de PInstitut Pasteur, April 25, 1897, Tome xi. No. 4, 
p. 289. — 10. Stokes and Wegefarth. Johns Hopkins Hosp. 
Bulletin, No. 81, Dec. 1897. — 11. Van der Loeff. Monats- 
hefte f. praktische Dennatologie, 1887, vol. vi. No. 5, p. 189. — 
12. Von Sicherer. Miinchener med. Wochenschrift, No. 34, 
1895, P- 492. 



Professor Michael Foster suggested to me some 
years ago that in view of possible deterioration of lymph 
stocks, after employment through a long series of 
generations, it would be well if possible to devise some 
practical means of investigating the efficiency of the 
protection afforded by some of those at present in use 
against variolous infection. 

The inoculation of smallpox on the human subject 
being now a penal offence, I turned my attention to 
the monkey tribe, on account of their similarity in 
many respects to man, although assured on high 
authority that they were not susceptible to either 
vaccinia or variola. On putting the matter to the 
test, however, I was agreeably surprised to find that 
this was not the case, inoculations of vaccine and 
variolous lymph having each of them given, in my 
hands, successful results in every instance in which I 
have tried them on the monkey. 

Briefly described, the modus operandi has been as 
follows : — The upper arm, or occasionally the inner 
surface of the thigh, is shaved on both sides of the 


body ; the skin is then washed with a solution of some 
disinfectant, such as corrosive sublimate or carbolic 
acid, and afterwards with alcohol. Each arm (or 
thigh) is then scarified in three or four places and the 
lymph well rubbed in. In the first series of experi- 
ments I used ether as an anaesthetic, but I after- 
wards found this quite unnecessary, as the monkey 
appears by no means to dislike the scratching process, 
becoming absolutely quiet as soon as the operation is 
commenced, and remaining with limbs limp and eyes 
half closed, as if hypnotised, until its termination, when 
it at once becomes as lively as ever. In the case 
alike of variola and of vaccinia the local result of 
inoculation attains its acme (qua vesiculation) in the 
monkey, as in the human being, about the eighth day. 

The first signs of reaction appear usually on the third 
day, by which time, if variolous lymph has been used, 
there is a distinct though very thin crust over the site of 
inoculation. In the case of vaccinia the appearance of 
crust at this date is less marked. By the fifth day 
vesiculation in both cases has generally commenced, 
this becoming more obvious up to the eighth day, 
though even then it is much less marked in variolous 
cases than in those which have been vaccinated, the 
difference being recognised with the greatest ease. 
Later, the vesicle gives place in either instance to a 
pustule, by which time there is not unfrequently con- 
siderable swelling of the skin and subcutaneous tissue 
and of the nearest lymphatic glands; the pustule 
gradually dries up, and a scab is formed, which is more 


1 30 VA CCINA TION : chap. 

pronounced in the case of vaccination than variolation, 
and which falls ofif some time during the third week, if 
the monkey have not picked it off before. 

The chief difference noted between the effects 
resulting from the local inoculation of these two 
diseases is, that in the case of variola there is more 
or less of a crust from the first ; that vesiculation is 
less marked than in vaccination ; that about the ninth 
to the eleventh day a general eruption may appear, 
which in some instances, as in a monkey which, at 
the request of Professor Foster, was inoculated at 
Cambridge, covers the whole surface of the body ; 
and that the final scab at the site of inoculation is 
not so elevated in the variolated as in the vaccinated 
animal. In both cases there is usually a rise of body 
temperature, which is more marked and longer sustained 
in variola than in vaccinia. After variolation, it was 
noticed in several cases that the monkey suffered from 
diarrhoea, that its eyes were suffused, and that it was not 
as active as usual. A peculiar odour was also noticed, 
quite distinct from the well-known smell of " monkey." 
In no instance had the experiment a fatal termination. 

Having thus proved to my own satisfaction that 
monkeys are susceptible not only to vaccination but 
also to smallpox, I next determined to make trial as to 
the protection against smallpox afforded in the monkey 
by previous vaccination, and the protection against 
vaccination afforded it by variolation, and I went on to 
compare the effect produced by the use of human and 
of calf vaccine respectively. 


For this purpose humanised lymph was obtained 
from Birmingham, where, to the best of my behef, the 
same strain has been continuously carried on by means 
of arm-to-arm vaccination for the past thirty-eight years. 
The calf lymph used was obtained from the Govern- 
ment Animal Vaccination Station in Lamb's Conduit 
Street, while smallpox lymph was supplied to me from 
the Hospital Ships, from Warrington, and from Man- 
chester. The following experiments will serve as illus- 
tration of the system I pursued : — 

Experiment i. — On 13th July 1892 I vaccinated a 
male rhaesus monkey in four places on the left arm 
with calf lymph, on points taken at the Animal Vac- 
cine Estabhshment on the previous day. On 20th 
July, the eighth day, the appearance presented was 
undistinguishable from that of a successful case of 
human vaccination. On 23rd July the vesicles had 
given place to four large scabs, raised nearly one-eighth 
of an inch above the surface of the arm, while the 
surrounding inflammatory zone had nearly disappeared \ 
the axillary glands on the same side were considerably 
enlarged. A drawing was made of the arm as it appeared 
on this day by Mr. Lapidge. By 9th August the scabs 
had all come off, and the monkey was again vaccinated, 
but this time on the opposite arm in two places, as 
well as on the inside of the right thigh also in two 
places, with humanised lymph of ascertained activity. 
The animal was carefully examined day by day up to 
17th August, but no sign of the second vaccination 
having " taken " was observed. 

132 VA CCINA TION : chap. 

Experiment 2. — On 19th July a large female rhaesus 
monkey was inoculated by scarification in eight places 
(four on each arm) with two tubes of smallpox lymph 
from the Hospital Ships. The lymph was obtained from 
a patient aged nineteen, stated to have been vaccinated 
in infancy, who was suffering from a semi-confluent 
attack of the disease. The lymph had been sealed up 
in carefully sterilised tubes. On 23rd July (the fifth 
day), the right arm of the monkey showed papulation 
and commencing vesiculation. The axillary glands 
were slightly enlarged. 

On 1 8th August, a month after inoculation, this 
monkey was vaccinated with calf lymph of known 
activity. No result of any kind followed this second 

It is, I think, hardly necessary here to enter into 
details of the numerous experiments which have been 
performed at intervals of one to five months after the 
primary inoculation, whether vaccinal or variolous, and 
I will merely add that in no instance did anything in 
the nature of a successful result follow the first or 
subsequent revaccinations. From these experiments it 
would appear that the protective power of lymph obtained 
from these three different sources, when inoculated on 
the monkey, is practically identical in all respects. In 
discussing the origin of the various lymph stocks at 
present in use, Messrs. Collins and Picton, in the Minority 
Report appended to the Final Report of the Royal 
Commission on Vaccination, make a point of the 
impossibility of employing at the present day what 


used to be known as the "variolous test," as a proof of 
the efficacy or the reverse of any particular strain of 
lymph. In view, however, of the results of my experi- 
ments with monkeys, this criticism of Messrs. Collins 
and Picton falls to the ground ; since if it is desired to 
apply the " variolous test " to any given lymph stock, 
all that is necessary is to vaccinate a monkey with 
a sample of the lymph in question, and subsequently 
to inoculate the animal with potent smallpox lymph, 
after the lapse of such period from the first operation 
as may be thought desirable. 


I. BuiST. Vaccinia and Variola, 1886, p. 84 et seq. — 
2. CoPEMAN. Proceedings of Royal Society, 1893, vol. liv. p. 
187 ; Journal of Pathology and Bacteriology, May 1894, p. 407 ; 
Transactions of Epidemiological Society, N.S. 1892-93, vol. xii. 
p. 142. 



In 1 89 1 I placed on record, in a paper presented to 
the International Congress of Hygiene, certain observa- 
tions relating to the effects of vaccination with calf 
lymph containing no living micro-organisms. As was 
to be expected, vaccination carried out with lymph, 
previously sterilised by exposure to heat, was not 
followed by the usual sequence of papule, vesicle, etc., 
arising at the site of insertion. Nevertheless, the 
calf which had been operated on by this apparently 
inert lymph was subsequently found to have been 
rendered more or less immune against re- vaccination 
with lymph of normal potency. 

My attention was first called to the possibility of 
thus obtaining temporary protection, at any rate, without 
production of local result, in the course of experiments 
carried out with lymph which had been sterilised by 
heat at a temperature of 60° C. Such lymph not only 
gave rise to no bacterial colonies in plate cultivations 
seeded from it, but also, as stated, it produced no local 
effect whatever when calves were vaccinated with it. I 
further found that, as in one experiment to which refer- 


ence may be made, exposure of the lymph to a temper- 
ature as low as 48° C. for an hour had been sufficient 
to so sterilise the lymph that when inoculated on a calf 
no apparent result was produced. A week later this 
calf was inoculated by Dr. Cory directly from another 
animal, the lymph from which was at the same time 
used with perfect success on other calves and a number 
of children. On this particular calf, however, the oper- 
ation performed by Dr. Cory was altogether without 
result, thus apparently showing that the use of lymph 
treated in this manner by exposure to a temperature 
considerably above that of the body, though itself 
producing no vesiculation, had rendered the animal 
insusceptible to the subsequent operation with fresh 
lymph of normal potency. 

On the first occasion on which these results were 
obtained, I thought that by chance the particular 
calf might have been the subject of an unusual 
insusceptibility, but results of later experiments show 
that this was not the case, since under similar cir- 
cumstances the result of the second vaccination was 
invariably negative. In consequence, no doubt remained 
in my mind that something of the nature of protection 
had been really afforded by the primary operation in 
each instance, although nothing in the way of local 
reaction had been perceptible. A similar course of 
events has been observed by Chauveau, Klein, and 
myself, in the conduct of variolation experiments on the 
calf. These experiments seemed to afford some evidence 
as to the possibility of obtaining all the desired effects 

136 VACCINATION: chap. 

of vaccination without likelihood of concomitant 
danger ; and I was, therefore, led further to experiment 
as to the possibility of obtaining similar immunity by 
the method of subcutaneous injection, which for certain 
reasons I considered would be preferable to that of 
multiple superficial incisions on the skin. And as the 
result of experiment in this direction, by the introduction 
into the subcutaneous tissue of the back of the monkey 
of vaccine lymph which had been previously diluted 
with normal saline solution, in order to have at disposal 
a quantity of fluid easier of manipulation, I found that 
some degree of protection was obtainable against sub- 
sequent vaccination carried out in the usual manner. 
In one of the two instances in which I employed this 
method, a local tumour, possibly an abscess, resulted, 
probably due to irritation from septic organisms or their 
products in the lymph. 

Chauveau records having pursued a similar method 
of subcutaneous inoculation of vaccine lymph on bovines 
and horses ; and confirms me in the opinion that by 
means of a generalised influence over the system, 
although unaccompanied by production of local result, 
it is possible to obtain (for a time at least) immunity 
from subsequent vaccination. 

With the hope of throwing some light on this pro- 
blem, and at the same time making trial of the value of 
what might possibly afford a new method of protection 
against smallpox, I devised the following experiment on 
somewhat similar lines to the well-known work of Tizzoni 
and Cantani on Tetanus. In a monkey which had 


been successfully inoculated with variola five months 
previously, and which, after an interval of three months 
had again been variolated and also vaccinated without 
result, the carotid artery was dissected out and tied with 
antiseptic precautions, after etherisation of the animal. 
A glass cannula was then inserted into the artery below 
the ligature and about 30 c.c. of blood received into 
two test-tubes, each of which contained a few drops of 
oxalate of potassium solution. The blood, which was 
thus prevented from coagulating, was then centri- 
fugalised, and rather more than half the total amount 
of transparent, slightly yellow-tinted plasma was obtained, 
the red corpuscles remaining in a dense mass at the 
bottom of the tubes. Of this plasma about 8 c.c. 
were then carefully inserted into the peritoneal cavity of 
a second monkey through a minute incision into the 
linea alba. Two days after the operation both animals 
appeared perfectly well. 

After an interval of fourteen days, on yth April 1893, 
the second monkey was vaccinated on the left arm, and 
also on the inner aspect of the right thigh, with calf 
lymph on points obtained from the Animal Vaccine 

I had no opportunity of seeing the animal again till 
the eighth day, owing to compulsory absence from town ; 
but it was then evident that complete protection, at any 
rate, had certainly not been afforded by the operation. 
All the scarified places had "taken," although in each 
instance the result produced was a somewhat abortive 
one, as was noticeable on comparing this animal with 


another which had been vaccinated on the same day 
and with the same lymph, and which therefore served 
as an excellent "control." 

Some effect had therefore apparently been produced 
on the system of the monkey by the procedure adopted, 
but at the same time it was obviously but slight, even 
though the amount of plasma injected was considerable, 
in relation to the size of the animal. 

In like manner, the results of experiments in this 
direction recorded by different observers are all some- 
what unsatisfactory as regards the production of a 
potent antitoxic serum. Thus Kramer and Boyce were 
unable to produce any appreciable immunity even with 
large doses of serum separated from the blood of 
previously vaccinated calves ; and, more recently, 
Beumer and Peiper have arrived at a similar con- 
clusion. On the other hand, Kinyoun, Hlava, and 
Houl claim successful results ; but, so far as can be 
judged from their publications, such claim would appear 
to rest on somewhat slender foundation. It may, 
however, be mentioned that Hlava and Houl record an 
instance in which injection of their serum to the extent 
of 0.6 to i.o C.C. per kilo of body weight of the experi- 
mental calf entirely prevented the action of potent lymph 
with which this calf was vaccinated four days later by 
means of superficial incisions on the skin. 

The fullest and most recent contribution to this 
subject is that of Becl^re, Chambon, and Menard. The 
conclusion at which these authors arrive, as the result 
of a long series of experiments, is that the serum of 


a previously vaccinated heifer, removed after the 
period of maturation of the vesicles, is capable, on 
injection into another animal, of conferring a certain 
immunity against the effects of subsequent vaccination ; 
but the diagrams with which the paper is illustrated 
show that the antagonism is by no means complete. 

In order to obtain the best result, these observers 
found it desirable to collect the serum at a period of 
from ten to fifteen days after vaccination of the calf, at 
which date they found it possessed -a maximum degree 
of potency, as judged by the effects of subsequent 
vaccination. They state also that the immunising 
action of the serum obtained from a vaccinated calf is 
very rapid, since a subcutaneous injection of the serum 
in sufficient amount, performed on a calf immediately 
before vaccination of the same animal by means of 
numerous insertions on the skin, was found capable of 
modifying the results of this after-vaccination to as great 
an extent as was apparent in any of their experiments. 

When, however, instead of employing for sub- 
cutaneous injection the serum from a previously 
vaccinated calf, they used vaccine lymph in similar 
fashion, immediately before vaccination of the same 
animal, the development of the resulting eruption did 
not appear to be in any way modified. But when 
vaccination was postponed till the eighth day after sub- 
cutaneous injection of vaccine lymph, a somewhat simi- 
lar result was obtained as in those instances in which, 
immediately before vaccination, the serum obtained 
from a previously vaccinated calf had been injected. 


The amount of serum which they found it necessary to 
employ in order to obtain even the incomplete degree 
of immunity recorded by them, was about looth the 
weight of the animal operated on. They further found 
that the immunising action of their serum was capable 
of demonstration, to some extent, even when employed 
for injection at an interval of twenty-four or even forty- 
eight hours subsequent to vaccination of the animal. 
Encouraged by the result of their experiments, M. Beclere 
employed some of the serum obtained from one of their 
experimental animals for the treatment of a case of 
smallpox. The amount of serum injected in this case — 
that of a woman aged twenty-nine years, and weighing 
78 kilogrammes — was a litre and a half, given in three 
successive injections during the course of an hour. The 
injection of this enormous quantity of serum appeared 
to cause no local or general injury, and the woman 
recovered rapidly and perfectly. M. Beclere, indeed, 
expresses the opinion that the employment of the serum 
exerted a marked influence in bringing about so success- 
ful a result. In his paper he refers to certain other 
cases in which a similar line of treatment had been 
attempted, which have been put on record by Dr. 
Auch^ of Bordeaux, and Dr. Landmann of Frankfort. 
These observers, however, employed in the treatment of 
their cases serum obtained from human beings immun- 
ised by a previous attack of smallpox ; but the amount 
of blood withdrawn afforded so small a quantity of 
serum that it is perhaps not matter of wonder that no 
appreciable result was found to follow on its use. 


About the same time Landmann carried out two 
further attempts at a similar method of treatment, 
employing in these instances, however, serum from a 
previously vaccinated heifer. Dr. Kinyoun and Dr. 
MacElliot also experimented in this direction in 
America; but, as in the case of the employment of 
serum from human beings, the amount of material em- 
ployed in each instance was again obviously too small to 
render it Hkely that any positive result could be expected. 

As far as I am aware, M. Becl^re and Dr. Kin- 
youn's cases are the only ones in which successful 
results are stated to have followed the attempt at treat- 
ment of human variola by the antitoxic method. And 
it would seem probable that no really useful results are 
likely of accomplishment until we are in possession of 
some more satisfactory method of immunising the 
system of the animal from which the serum is derived. 
Such a consummation can only be expected when 
further research shall have provided us with reliable 
methods for the ready cultivation, outside the animal 
body, of the microbe specific to variola. 


Antitoxin : i. BECLfeRE, Chambon, and Menard. Annates 
de rinstitut Pasteur, and Bulletin de la societe mddicale des 
Mpitaux, Jan. 1896. — 2. Beumer and Peiper. Berl. klin. 
Woch. August 26, 1895. — 3. Hlava and Houl. Wien. klin. 
Rundschau, Oziohox (i2,xiA 13,1895. — 4. KiNYOUN. Philadelphia 
Med. News, Feb. 2, 1895. — 5. Kramer and Bovce. Brit. 
Med. Journal, 1893, vol. ii. p. 989. — 6. Landmann. Zeitschrift 
fiir Hygiene, 1895, vol. xviii. — 7. Raynaud, Maurice. Comptes- 
rendus, 1877, vol. Ixxxiv. p, 453. — 8. Sternberg and Reed. 
Trans. Assoc. American Physicians, Philadelphia, 1895, PP* 57-69- 



By the term " Animal Vaccination," as it is now applied, 
is meant the employment for the vaccination of the 
human subject of the virus of cow-pox, as propagated 
upon a succession of calves or heifers, the original virus 
having been derived from pocks upon the cow spon- 
taneously affected with the natural disease. 

The late Dr. Ballard, in his prize essay on vaccination, 
gives a most interesting and valuable historical Account of 
the introduction of the practice. In this essay he states 
that the practice of animal vaccination appears to have 
originated with M. Negri in Naples, who was seemingly 
the first to propagate natural cow-pox by successive 
inoculations upon the heifer. Prior to his time, says 
Dr. Ballard, cows had been, in various places and by 
various persons, inoculated with vaccine virus derived 
from the human subject, the lymph obtained from pocks 
thus produced being used for human vaccinations. 
This practice has been termed " retro -vaccination." 
There appears to be no great difficulty or uncertainty 
attending retro -vaccination, so long as animals suffi- 
ciently young are made its subjects, and the human 


lymph is taken under circumstances favourable to its 
activity. The general result of reinoculation of it upon 
the human subject has been that the character and 
activity of the virus thus retransferred remained 
unaffected by its transmission through the system of the 
cow. This was the practice first pursued at Naples by 
Troja in 1805, shortly after the introduction of vaccina- 
tion into Italy, and also pursued by his successor 
Galbiati. When, however, M. Negri, in 1842, suc- 
ceeded to the practice on the death of Galbiati, he soon 
gave up retro-vaccination, and began to propagate the 
virus by inoculation from one cow to another, that first 
propagated being of human origin. Subsequently, on 
the occasion of an outbreak of cow-pox in Calabria, the 
use of this virus was abandoned, and M. Negri propa- 
gated thenceforth that obtained from the natural cow- 
pox. It is said that on two occasions he thus availed 
himself of natural cow-pox matter obtained in Italy, but 
that on a third occasion, in 1858, on which he renewed 
his supply, it was derived from London. 

A good deal of doubt has been thrown upon the 
origin of this third renewal of virus, and it has been 
questioned whether it was virus from natural cow-pox 
at all. However, it appears that it is this virus which 
M. Negri propagated subsequently to 1858. In 1864 
M. Lanoix, a young French physician, visited Naples 
in order to study the practice of animal vaccination as 
pursued there. On his return, he brought back with 
him to Paris a calf inoculated with the virus then in 
process of transmission by M. Negri, and in concert 

144 VACCINATION : chap. 

with M. Chambon set up a private establishment for 
the propagation of the virus from calf to calf, and for 
the supply of animal vaccine in Paris. 

In his report on vaccination in France during 1864, 
M. Depaul in the name of the Academie de Medecine 
gave an account of the observations made in Paris on this 
subject, and in February 1866, the French Government 
placed a sum of 6 000 francs at the disposal of the Academic 
for the purpose of experiment in the matter. A Com- 
mission was appointed, the report of which was drawn 
up by M. Depaul. This report was favourable to the 
practice of animal vaccination. The Commission, how- 
ever, were much divided in opinion, and the Academie 
did not then recommend that the practice should be 
adopted. At first, the virus used by the Commission 
was that in use by MM. Lanoix and Chambon, and 
which they had previously obtained from Naples. But 
after four transmissions of this lymph had been made, a 
new source of virus from natural cow-pox was discovered 
at Beaugency (Loiret), and then the use of the 
Neapolitan strain was abandoned and this new and 
undoubted cow-pox virus alone employed. MM. Lanoix 
and Chambon also adopted the use of this strain to 
the exclusion of that they had before employed, and 
having, in the autumn of the same year (1866), met with 
another case of natural cow-pox at St. Mande, near 
Paris, they introduced this lymph also into their practice 
of animal vaccination. They saw no advantage in 
keeping these two lymphs distinct, and the strain which 
they have employed from 1866 onwards is a mixture of 


the two natural sources discovered at Beaugeilcy and at 
St. Mande. 

Whatever the origin of the NeapoUtan virus may 
have been, therefore, there can be no question that the 
inoculations made by the Commission (after the first 
four), and those afterwards made in Paris by MM. 
Lanoix and Chambon, were transmissions of the virus 
of a cow-pox of spontaneous origin. 

From Paris the practice of animal vaccination ex- 
tended in many directions. Thus in February 1865, 
it was introduced into Brussels by Dr. Warlomont, 
whose first supply of lymph was obtained from M. 
Lanoix (Neapolitan source). At a later period, he 
obtained from Paris virus in course of propagation from 
the Beaugency source, and, still later on, he introduced 
a third source obtained in July 1868 at Esneux (Liege). 

In September 1866, at the request of the Minister 
of the Interior, the Royal Academy of Belgium referred 
the whole question to a commission consisting of MM. 
Bellefroid, Thieruesse, and Marinas, who reported 
favourably upon it, and, in consequence, in July 1868, 
an " Institut Vaccinal de I'Etat " was estabUshed at 
Brussels, under the direction of Dr. Warlomont, for 
maintaining the practice of animal vaccination, and 
distributing gratuitously lymph obtained from the 
animals, or lymph taken from children who had been 
vaccinated direct from the animals, according to the 
preference of the vaccinators or other practitioners 
demanding it. 

The practice was introduced into Berlin by Dr. 



Pissin in June 1865, and the establishment which he 
founded is still carried on as a private speculation. 
Animal vaccination appears to have been introduced 
into Vienna in a similar manner about the same time, 
and many of the great capitals of Europe rapidly 
followed suit. Thus, early in 1868, the Society for the 
Promotion of Cow-pox Inoculation at Rotterdam deter- 
mined to open a station for the practice of animal 
vaccination, and they commenced operations in April 
of that year, distributing lymph throughout Holland 
and its colonies. A similar course was adopted in the 
following year by the corresponding society of Amsterdam. 
Switzerland also owes the comparatively recent intro- 
duction of animal vaccination into that country to 
private enterprise ; what is now known as the Institut 
Vaccinal Suisse having been founded in 1882 by M. 
Charles Haccius, the present Director of the establish- 
ment. Originally a private venture, the Institute, which 
is situated at Lancy on the outskirts of Geneva, is now 
recognised by the various cantonal governments, 
M. Haccius, in consideration of an annual subvention, 
supplying to public vaccinators throughout Switzerland, 
free of cost, all the lymph required by them in the 
performance of their duties. This estabhshment I have 
recently (1897) had the opportunity of visiting, in the 
course of a tour of inspection, at the instance of the 
Government, of certain of the chief calf-vaccine estabUsh- 
ments on the Continent of Europe. 

At Paris, as is the case with the establishment of 
M. Haccius at Lancy, the Institut de Vaccine Animale 


is practically a private concern, although the muni- 
cipality of the city of Paris contract with the Directors 
to carry out all such public vaccinations, within their 
jurisdiction, as may be necessary. This institution, 
which is now carried on by M. Chambon and Dr. St. 
Yves Menard, was founded, as already mentioned, by 
M. Lanoix. Originally situated in the Rue Massillon, 
it was subsequently transferred to the Rue Ballu, where 
what was formerly a dwelling-house has been adapted 
to its present purpose. 

In Germany all the establishments visited by Sir 
Richard Thorne and myself, at Berlin, Dresden, and 
Cologne respectively, are under Government control. 
Of these, that at Cologne is the most recent of its kind, 
and its buildings with their fittings are of the most 
modern description. This Institution, of which Dr. 
Vanselow is the Director, was erected in 1889 for the 
production of animal lymph, to meet the needs of the 
Rhine province and Hohenzollern districts. The build- 
ing adjoins the central meat market, a matter of con- 
venience for the obtaining and subsequent disposal of 
calves. On the ground floor of the building are the 
collecting room, calf stable, slaughter room, director's 
and clerk's rooms, corridor, and offices, while on the 
first floor is a bacteriological laboratory, fitted up with 
all needful apparatus, which is reached from the collect- 
ing room by an iron spiral staircase. 

At Brussels, the propagation, storage, and distribu- 
tion of calf lymph is carried out at the Ecole 
Veterinaire, under the supervision of Professor Degive, 


the Director of the establishment. The accommoda- 
tion provided in the present building is regarded as 
very insufficient for the purpose, and accordingly, as 
Professor Degive informed us, a new vaccinal institute 
is about to be erected, in the construction of which 
all modern scientific requirements will be met. The 
private establishment, originally founded by Dr. Warlo- 
mont, and which is now carried on by his son-in-law, 
an officer in the Belgian army, we had not the oppor- 
tunity of visiting. 

Although the practice of animal vaccination is now 
practically universal throughout Continental Europe, in 
England up to the present time it has been employed 
to a very limited extent. At the Government Animal 
Vaccine Establishment at Lamb's Conduit Street, 
which was founded in 1881, as the result of investiga- 
tions by the late Sir George (then Dr.) Buchanan and 
Dr. Cory, direct vaccinations of children, from calf to 
arm, have been carried out from that date onwards to 
the present time. 

From the calves vaccinated at Lamb's Conduit 
Street, lymph has also been supplied to public vaccina- 
tors in quantity sufficient to enable them to start their 
periodical vaccinations, and provision has also been 
made for supplying a comparatively small amount of 
lymph to private practitioners in addition. The lymph 
thus sent out has for the most part been preserved by 
drying on ivory " points," although a certain small 
proportion has been stored in the Hquid form in 
capillary glass tubes. 


Until quite recently, however, all public vaccina- 
tions in this country, with the few exceptions just 
mentioned, were required to be performed by direct 
transmission of the virus from arm to arm, stational 
attendances at fixed periods having been arranged for 
in order to render feasible such method of operating ; 
weekly attendance having been provided in large towns, 
while quarterly or even half-yearly periods sufficed for 
the required purpose in scattered country districts. 

As the result, however, of the findings of the Royal 
Commission on Vaccination, legislative measures have 
now been introduced by which arm-to-arm vaccination 
will be entirely superseded, in public work at any rate, 
by animal vaccination ; and the adoption of the use of 
preserved glycerinated calf lymph having been de- 
termined on, the necessity for stational attendance will 
disappear, except in so far as may appear desirable for 
purposes of control or in order to provide facilities for 

The principal grounds on which the practice of 
animal vaccination has been advocated are : 

I. The opinion, widely though not universally held, 
that the results of vaccination from arm to arm are not 
such as they used to be in .the earlier years of the prac- 
tice of vaccination ; in fact that by repeated human trans- 
mission the virus has become weakened, and that the 
pocks produced by the introduction of lymph which has 
passed through a large number of human beings are not 
so fine or so perfect as those which result from the use 
of animal lymph. Jenner himself indeed appears to have 

I50 VACCINATION : chap. 

been under the apprehension that lymph-stocks would 
be liable to degenerate as the result of long-continued 
transmission through the human subject. 

In the present state of our knowledge, however, such 
enfeeblement of the specific virus can hardly be regarded 
as probable, except under conditions that may be 
obviated by reasonable skill and care on the part of the 

Jenner early discovered that vaccine lymph only 
exhibited its full degree of activity when taken at the 
stage of maturation of the vesicle, and before its con- 
tents became at all purulent. If this precaution be 
observed, together with strict cleanliness in removal and 
insertion of the lymph, experience has shown that no 
appreciable degeneration can be demonstrated. 

2. The opinion that certain other human diseases 
may be propagated together with vaccinia, when 
vaccinations are performed from arm to arm. Galbiati, 
who was one of the first, if not the first, to vaccinate 
with bovine lymph, adduced this belief as his reason for 
practising vaccination from the cow; arguing that on 
vaccinating the cow from the human subject vaccinia 
alone would be communicated to her, any other morbid 
germs which might be introduced along with it remain- 
ing without effect. He considered apparently that in 
practising retro-vaccination according to the method 
which he devised, he, as it were, filtered human vaccine 
material from any possible contamination. 

As the result of all the evidence brought before 
them during the lengthy period of seven years, the 


English Royal Commission have reported that the risks 
incurred from arm-to-arm vaccination " are undoubtedly 
real and not inconsiderable in gross amount," although 
" when considered in relation to the extent of vaccina- 
tion work done they are insignificant." And they add 
that " there is reason further to believe that they are 
diminishing under the better precautions of the present 
day, and with the addition of the further precautions 
which experience suggests will do so still more in the 

It is indeed the fact that cases of syphilis have 
been occasionally conveyed from one human being to 
another as the direct result of vaccination ; and although 
the number of cases that can be authenticated is 
extraordinarily small in number, there is advantage in 
the adoption of a method which, while equally 
efficacious, entirely obviates the possibility of so lament- 
able an occurrence. 

Erysipelas and tuberculosis are also two of the not 
inconsiderable number of diseases with the spread of 
which vaccination has been credited. No doubt the 
greater proportion of cases of erysipelas following on 
the process are of what has been termed " late " origin. 
These are due to infection of the vaccination wound at 
or about the time when it has been customary to open 
the vesicle. The general use of animal vaccine, by 
obviating the necessity for opening the vesicle, may be 
expected to be followed by a diminution of cases 
originating in the manner suggested, though accidental 
injury to the arm, or the use of improper dressings, will 

152 VACCINATION chap, viii 

doubtless prevent the complete disappearance of erysipe- 
las or other septic diseases. Invaccinated erysipelas 
however, we may anticipate, will shortly become in 
great measure, at any rate, a thing of the past ; and, 
fortunately, the possibility even of the invaccination of 
tubercle may be eUminated as the result of the treatment 
of calf vaccine with glycerine as now officially adopted 
by the Government, after a method which I first 
advocated some years ago, and to which I at once 
proceed more particularly to refer. 


I. Ballard. •' Animal Vaccination," Report to Privy Council, 
1868. — 2. BousQUET. Gazette midicale, 1866, p. 319. — 3. 
Marinus. Bulletin de Vacad. roy. de mddecive de Belgique, ser. 
ii. torn. ix. No, 8. — 4. Murphy. "Animal Vaccine Establish- 
ments of Holland and Belgium," i?^/£>r/ of Med. Officer to Local 
Govermnent Board, 1883, p. 27. — 5. Sanderson, Burdon. 
Sixth Report of Med. Officer to Privy Council, 1 863, p. lo. — 6. 
Seaton. Twelfth Report of Med. Officer to Privy Council, 1869, 
p. 171. — 7. Report of Med. Officer to Local Government Board, 
1878, p. 7. 

The Filial Report of the Royal Commission on Vaccination may 
also be consulted with advantage. 



In a paper presented to the International Congress 
of Hygiene held in London in 1891, and subsequently 
published in the Transactions of that Congress, I called 
attention to a special method for the bacteriological 
purifi cation and preservation of vaccine lymph. This 
method consisted in the intimate admixture of a given 
amount of lymph, or rather vesicle pulp, with a sterilised 
50 per cent solution of chemically pure glycerine in 
distilled water, and in subsequent storage of the result- 
ant emulsion, in sealed capillary tubes, for several weeks. 
For some years antecedent to 1891, I had been 
engaged in investigating the nature and mode of action 
of the specific virus contained in vaccine lymph, chiefly 
from the bacteriological point of view. Early in the 
course of my experiments it struck me that the exuberant 
growth commonly manifested by what were so evidently 
" extraneous " organisms, might be very possibly inter- 
fering with, if not superseding, the more important and 
essential organism that I was seeking. I therefore 
turned my attention to the discovery, if possible, of 
some means of so treating vaccine lymph as to inhibit 

154 VACCINATION : chap. 

the multiplication in it of all " extraneous " organisms, 
and eventually to destroj^ them altogether, without at 
the same time injuring its potency for vaccination. 
These bacteria evidently find in the lymph, especially 
when removed from the body, a suitable medium for 
their subsequent multiplication, while at the same time 
it would appear that growth and multipHcation of them 
has as result the gradual inhibition of the specific effect 
proper to the vaccine virus itself Upon all grounds, 
then, the obvious indication for my guidance w^as not 
only to prevent such multiplication of " extraneous " 
organisms subsequent to storage in the usual manner, 
but, if possible, to remove them altogether as soon as 
the lymph was taken from the living subject, without 
injury to the actual contagium of vaccinia. 

Without detailing the various experiments adopted 
with this object in view, it is necessary here to state briefly 
the lines on which the work was carried out. In the 
first place, trial w^as made of the method of exposure of 
lymph for definite periods to a temperature consider- 
ably above blood-heat, which had, in the hands of 
Kitasato, met with conspicuous success in the isolation 
of the bacillus of tetanus. Proceeding in this manner, 
and in every experiment observing the precaution of 
making control cultures, I presently arrived at a tem- 
perature, exposure to which is apparently incompatible 
with the continued existence of those micro-organisms 
which can ordinarily be grown when vaccine lymph is 
inoculated into nutrient jelly. The temperature in 
question ranged between 38° C. and 42° C, but the 


method did not in practice prove advantageous. Thus, 
if plate cultures were made of lymph after exposure for 
an hour at the lower register, a few points of growth 
occasionally after the lapse of a day or so made their 
appearance ; and, on the other hand, the higher tem- 
perature, though it inhibited all extraneous microbes, 
appeared sometimes to exert an injurious effect on the 
lymph, in so far as regards the normal vesiculation which 
should result from its inoculation in the living animal. 
Some method of readier application, and requiring less 
delicate manipulation, was therefore obviously desirable. 
This I at length found in the addition to the lymph, or 
rather to the vesicular pulp, obtained from a vaccinated 
calf, of a sterilised 50 per cent solution of glycerine 
in distilled water. 

Admixture of glycerine and vaccine lymph is, of 
course, no new device. Indeed, my attention has 
recently been directed to a letter by the late Mr. R. 
Cheyne, which appeared in the Medical Tunes as long 
ago as March 1850, in which he advocates the superiority 
of lymph which has been kept in the fluid state by 
addition of " the least portion " of glycerine over that 
which had been dried on points. As Mr. Cheyne's 
work appears to have been overlooked until quite 
recently, it may be of interest to set out in his own 
words his mode of procedure. " My method," he says, 
"is to take all the lymph I can get from the eighth day 
vesicle on a glass stopper (elongated in a narrow tongue- 
shaped form for about an inch below the neck of the 
small bottle into which it fits), and having accumulated 

156 VACCINATION: chap. 

it on both surfaces of the stopper near its end, I mix it 
well with the point of the probe with the glycerine held 
by the latter. Then after the stopper is put into the 
bottle (which is always to be kept standing up), the 
lymph, then assisted by gravity, will collect itself into a 
distinct drop on each surface of the stopper to be ready 
for use, as I know by experience, during a very long 
period, and in any number of cases, that would not 
exhaust the stock." In a further letter Mr. Cheyne 
acknowledges his indebtedness to the late Mr. J. 
Startin's advocacy of the therapeutic applications of 
glycerine, by which he learnt of the suitability of this 
substance for the object he had in view, and sets out, 
practically in the wording of Mr. Startin's paper, the 
distinctive properties of this substance. He goes on to 
say, " I mixed glycerine with vaccine lymph in the way 
detailed, .... with the satisfactory result, better than 
any theory, of discovering that, in addition to its known 
property of preventing fermentation and mouldiness in 
vegetable substances, it had also that of keeping vaccine 
lymph, an animal product, undecomposed in a fluid 
state for months." Three years later (in 1853), Mr. 
Cheyne showed to the Presidents of the Royal Colleges 
of Physicians and Surgeons a child whom he had 
successfully vaccinated with lymph which he had pre- 
viously kept for six months after treatment with 

It was left for Miiller of Berlin, in 1869, to demon- 
strate the further point that the quantity of material 
available could be increased by the addition of glycerine 


without injuriously affecting the potency of the mixture. 
He showed that vaccine lymph might be diluted with 
three times its bulk of glycerine, and still retain its 
properties unimpaired, a fact which has been taken 
advantage of at many of the continental vaccine stations, 
and by more than one purveyor of lymph. Cursch- 
mann, writing on the subject of smallpox in Zeimsenn's 
Encyclopaedia^ refers to this method as follows : — " Miiller 
has the great credit of having discovered the fact that 
by mixing vaccine matter with glycerine in certain pro- 
portions, the activity of the former is not diminished, so 
that we have here a means of increasing the volume of the 
lymph when the quantity is small or when there is an 
unusual demand for ^t. . . . The lymph and glycerine 
mixture appears to keep as well as the unmixed lymphs ^ 
But from this statement it is quite obvious that the sole 
object of employing glycerine in the manner described 
was to increase the amount of material available for 

Also with this end in view, glycerine was used by 
Dr. Stephen Mackenzie, at the London Hospital, 
in the epidemic of 1870-71, the mixture of lymph 
and glycerine being made up immediately before 
it was required for a large series of revaccinations. 
Similar means for increasing their amount of available 
lymph have been frequently employed by public 
vaccinators (among the earliest, my present colleague, 
Dr. Bruce Low) and others in times of stress. 

^ The italics are my own. — S. M. C. 


In 1882 Dr. Warlomont patented, in England, a 
method of admixture of glycerine with vaccine lymph, 
but no mention of the use of glycerine in this connection 
was made in the advertisements of his agents until 
within the last few months. Until recently, moreover, 
there would seem to have existed no appreciation of the 
inhibitory action exerted by the glycerine in bringing 
about bacteriological purification of the lymph, when 
the mixture is stored for some time prior to use, under 
conditions preventing access of air and light. 

When, however, a glycerine emulsion is properly pre- 
pared after the method I have advocated, it is found that 
the growth of extraneous aerobic bacteria is at once greatly 
inhibited, while after a longer or shorter period they are 
practically all killed out. This effect is best demon- 
strated by making a series of plate cultivations from 
tubes of glycerinated lymph, at gradually increasing inter- 
vals of time, a control plate being established in each 
instance from a specimen of the lymph material prior to 
the admixture of the glycerine. These observations of 
mine, since their publication in 1891, have received 
ample corroboration from a number of observers in 
various parts of the world, as follows : — 

In 1892, a paper dealing with this question was 
pubHshed by MM. Chambon and St. Yves Menard, in 
which they relate their experience of the use of 
glycerinated calf lymph when kept for a considerable 
period in capillary glass tubes (previously sterilised) 
closed by the blow -pipe. Not only were the results 
they obtained with originally active lymph highly satis- 


factory, but lymph which, in its fresh state, had given 
mediocre results, produced, after fifteen days' admixture 
with glycerine, a passable vesicle, and, after forty, fifty, 
or sixty days, a typical one. The improvement in the 
activity of such lymph seemed to them to be due to the 
gradual extinction of extraneous microbes under the 
combined influence of glycerine and length and method 
of storage. Professor Straus, who made plate cultures 
with their glycerinated lymph, found that, when fresh, 
it gave rise to numerous colonies of various microbes, 
especially staphylococcus pyogenes aureus and staphylo- 
coccus albus, but that when it had been stored from 
fifty to sixty days, plate -cultures therefrom remained 
absolutely sterile as regards these extraneous microbes. 
Samples tested at intervals between these two extremes 
presented fewer and fewer microbes as they became 
older. These experiments were repeated many times, 
and invariably with similar results. This evidence, so 
entirely corroborative of my own work, is the more 
important as it appears certain, from a perusal of their 
original paper, that the authors were ignorant that 
similar results had been previously arrived at, and that 
these had been published nearly twelve months before 
the appearance of their article. 

The value of glycerine in this connection is also 
strongly advocated by Leoni in a paper read before the 
Medical Congress held at Rome in April 1894, and 
afterwards pubHshed in the Revue d" Hygiene. He finds 
that vaccine lymph, as freshly collected, is apt to contain 
large numbers of micro-organisms, some of which, he 


says, are capable of exerting pathogenic properties when 
inoculated into the system along with the true vaccine 
virus. And he states that these microbes disappear 
completely from, or that at least their number is vastly 
decreased in, vaccine which, having been prepared with 
glycerine, is afterwards preserved for a period of from 
one to four months before use. His conclusions may 
perhaps be best given in his own words : — " Le vaccin 
r^cemment recueilli est un vaccin contamine." " Les 
agents de la contamination s'epuisent dans le vaccin 
conserve pendant quelque temps dans la glycerine." 
" Le vaccin conserve dans la glycerine pendant 1^4 
mois apres la recolte, represente le type du vaccin pur^ 
^MWQ virulence uniquement specifiquey "C'est de cette 
qualite de vaccin que I'hygieniste doit aujourd'hui tenir 
compte dans la prophylaxie de la variole." 

Dr. Klein also bears witness to the power exerted 
by glycerine in freeing vaccine lymph from bacteria. 
In speaking of the organism specific to vaccinia as 
being probably a spore-bearing bacillus, he says : " . . . 
it is established that the active principle of vaccine is 
preserved in glycerine, although, as is also known, 
glycerine acting for long times is a germicide for cocci 
and sporeless bacilli." 

In 1896, a Commission under the Presidency of 
Dr. Schmidtmann, and including Dr. Koch, Dr. Pfeiffer, 
and Dr. Frosch, together with the Directors of the 
Vaccine Institutes of BerHn, Cologne, and Stettin, was 
appointed by the German Government to inquire and 
report as to the best methods for the collection, preser- 


vation, storage, distribution, and use of vaccine lymph. 
In their report, which has been recently published, the 
Commission arrive at the conclusion, among others, 
that fresh lymph contains numerous microbes, the 
number of which on the addition of glycerine diminishes 
as the age of the mixture increases. To determine to 
what extent glycerine is efficacious in destroying the 
vitaHty of various A&'^xiiX.^y pathogenic microbes, numerous 
streptococci and diphtheria bacilli were mixed with 
specimens of lymph. As a result, the streptococci were 
killed in eleven days and the diphtheria bacilli in twenty 

Attempts by these experimenters, as also by Kitasato 
in Japan, to make other chemical agents serve for 
rendering vaccine lymph free from bacteria led to no 
results of value, as although the lymph could thus be 
rendered free from extraneous bacteria, it was found 
to be inefficacious as vaccine. Their next procedure 
was to determine the amount of glycerine that could be 
added to lymph, so as to exert a powerful action in 
purifying it from extraneous microbes, without in any 
way interfering with its specific action when employed 
for the vaccination of children or calves, and they came 
to the conclusion that the mixture of glycerine with 
distilled water could be employed to the extent of from 
fifteen to twenty times the weight of vesicle pulp 
collected, without interfering with the value of the 
material for the purpose of vaccination. 

The fact that the growth of the tubercle bacillus 
is encouraged in a nutritive medium containing about 


1 62 VA CCINA TION : chap. 

6 per cent of glycerine, in no way militates against our 
experience as to the germicidal value of this substance, 
when used in a strength of from 40 to 50 per cent. 
That certain drugs, when introduced in small quantities 
into the human economy, produce effects vastly 
different to those resulting from the introduction of the 
same drug in large quantities, is a fact familiar to every 
practitioner, and it is also well known to bacterio- 
logists that micro-organisms are profoundly affected by 
variations, in one or another direction, in the chemical 
composition of the particular medium in which they 
are placed. 

By consent, therefore, of numerous observers fully 
qualified to judge of the matter, we have in glycerinated 
calf lymph, properly prepared, a vaccine material which, 
while even more efficient as vaccine than the original, can be produced practically free from the 
extraneous organisms which, at one time or another, 
have been isolated from fresh or stored lymph by the 
method of plate cultivation. Of not least importance 
is the fact that, as shown by Dr. Blaxall and myself, 
vaccine lymph may by this method of preparation with 
glycerine be rendered free also from pathogenic bacteria, 
such as those of tubercle and erysipelas, even when 
these have previously been added in considerable 
quantity for experimental purposes. 

At a meeting of the British Association held at 
Liverpool in 1896, Dr. Blaxall and I communicated a 
report on the " Influence of Glycerine upon the growth 
of Bacteria/' 


In view of the publication of the Report of the 
Royal Commission on Vaccination, and of the reference 
to my previous work contained therein, it had appeared 
to us desirable to investigate more accurately the action 
of glycerine on various definite micro-organisms of a 
pathogenic or non-pathogenic nature respectively. 

For the purposes of this investigation quantities of 
glycerine were added to tubes of beef-peptone-broth, 
which were subsequently inoculated with equal quantities 
of pure cultivations, and incubated at blood-heat and at 
the room temperature respectively. Control inocula- 
tions in ordinary beef- broth were also invariably 
employed. Subsequently inoculations were made from 
the broth tubes on to solid media at varying intervals 
of time, in order to see whether the particular microbe 
still remained capable of growth or not. In all, some 
hundreds of inoculations were made, and the paper 
included a table in which were given the maximum 
limits of resistance attained in the different series. 

Practically, the results of each similar series were 
found to agree very closely. 

The micro-organisms employed for the inoculations 
comprised Staphylococcus pyogenes aureus, S. pyogenes 
albus. Streptococcus pyogenes, Bacillus pyocyaneus, 
B. subtilis, B. coli communis, B. diphtheriae, and B. 
tuberculosis. Smallpox and vaccine material in the 
form of " crusts " and lymph was also employed. 

Results. — I. No visible development of the micro- 
organisms employed took place in the presence of more 
than 30 per cent of glycerine. 


2. None of the micro-organisms experimented with 
could be recovered after exposure for a month to the 
action of from 30 to 40 per cent glycerine, with the 
exception of B. coli communis and B. subtilis when 
kept in the cold. 

3. B. coli communis, unlike B. typhosus, resists the 
action of 50 per cent glycerine in the cold for a con- 
siderable period, a fact likely to prove of value as an 
addition to our present methods of differentiating these 
microbes one from another. 

4. The samples of smallpox and vaccine material, 
whether as " crusts " or lymph, were sterilised com- 
pletely, so far as extraneous microbes are concerned, in 
a week, by the presence of glycerine to the extent of 
about 40 per cent in the broth -tubes. This short 
period of resistance is, doubtless, in part, to be explained 
by the fact that the smallpox crusts used in these 
experiments had been obtained several months before- 
hand. Presumably therefore the number of microbes, 
which had been able to survive for so long a period the 
process of drying, would be much less than might be 
expected to be present in " crusts " recently obtained. 

In this paper also, we published an account of some 
experiments relating to the action of glycerine on certain 
yeasts ; under this heading being included, for reasons 
of convenience, both Saccharomyces and Torulae. 

The results show that these organisms vary greatly 
in their resistance to glycerine, those not unfrequently 
found in vaccine material being easily killed out, whereas 
the common pink yeast (Rosa Hefe) resists the action 


of 50 per cent glycerine for an almost indefinite 

Since the date of the paper referred to, we have 
instituted a further series of experiments upon the 
tubercle bacillus, with the object, now, of determining 
whether this organism can survive, and remain capable 
of further development, after a sojourn for varying 
periods in glycerinated vaccine lymph. For this purpose 
vaccine material was rubbed up in the usual way with a 
steriHsed mixture of 50 per cent glycerine and water. 
The greater part of the resulting emulsion (containing 
glycerine to the extent of 42 per cent) was at once 
stored in small, 3-gramme, test-tubes. To the residue, 
amounting to about 4 c.c, was added a large quantity 
of growth from a culture of tubercle bacilli recently 
isolated and of full activity. This growth was thoroughly 
mixed with the emulsion, and the whole was poured into 
two I -gramme test-tubes which were corked and placed, 
with the rest of the tubed emulsion, in a cupboard in 
which our lymph is stored. This cupboard is kept at 
about 1 5° C. by means of ice. At the same time control 
inoculations were made from the tubercle culture in 
tubes of 6 per cent glycerine agar-agar, and in tubes of 
6 per cent glycerine-peptone-beef-broth (Roux's Media). 
These latter were then incubated at 37° C. 

At the end of a month the glycerinated vaccine 
emulsion was demonstrated, by means of plate-cultiva- 
tions, to be free from extraneous organisms; and, 
similarly, plates poured from the small tubes containing 
the tubercle bacilli also showed no growth. 

1 66 VA CCINA TION : chap. 

Inoculations were made from these small tubes on 
to tubes of 6 per cent glycerine agar-agar, and on to 
tubes of solidified blood serum. These were incubated 
at 37'' C. After a month's incubation, no growth 
resulted from any of these inoculations. Lest traces of 
glycerine carried over with the inoculation needle should 
have retarded or prevented the growth of the tubercle 
bacillus, some of the emulsion was freely mixed with 
sterile beef-broth, and from this also numerous inocula- 
tions were made and incubated at 37° C. These 
again, after the lapse of a month, failed to show any 
sign of growth. 

The control tubes inoculated with the growth of 
tubercle employed in the whole series of experiments 
and incubated at 37° C. all exhibited a copious growth 
in a month, and sub-cultures from them were all in turn 

As the result of our different series of experiments 
with the tubercle bacillus, then, it has been found 
impossible to recover this micro-organism after exposure 
for a month to the action of glycerine present to the 
extent of about 40 per cent, either in sterile beef-broth 
• or in fresh vaccine material. 

Valuable corroboration of our work in this direction 
is afforded by the results, as yet unpublished, of certain 
experiments carried out by Dr. Klein, which demonstrate 
the germicidal influence of glycerine on the tubercle 
bacillus. These I am now enabled by the courtesy of 
Dr. Klein to place on record. 

" \Qth February 1897. — From a typical tubercle 


culture put a good deal of growth into 50 per cent 
glycerine broth. After shaking, made two inoculations 
on slanting surface of glycerine agar; sealed and 
incubated at 37° C. . . . 

" 2 Ofth February, — Good growth of tubercle bacilU in 
both tubes. . . . 

" ^th April. — From glycerine - broth tubercle mix- 
ture of loth February, inoculated two glycerine -agar 
slanting surfaces; sealed and put in incubator at 37° 
C. . . . 

^'' 2 2,rd April. — No growth in any tube. . . . 

" 1st May. — No growth in any tube. . . . 

" ^Ih June. — No growth in any tube. ..." 

On 9th April 1897 Dr. Klein "injected two guinea- 
pigs subcutaneously with a good dose of the glycerine- 
broth tubercle mixture. There was no effect produced 
either locally or generally." 

Dr. Blaxall and I, feeling that cultivation experiments 
alone could not be held to be absolutely conclusive, 
have also carried out a series of injection experiments on 
guinea-pigs. The material we employed consisted of 
glycerinated vaccine lymph emulsion to which at the 
time of its preparation, at least a month before testing 
it by means of subcutaneous injection, virulent tubercle 
bacilH had been added in large quantities. For the 
purpose of these experiments two tubes, each containing 
three grammesof glycerinated calf lymph, were inoculated, 
immediately after preparation of the lymph, with three 
large platinum wire loopfuls, to each tube, of tubercle 
baciUi from a culture of proved activity. After securing 

i68 VACCINATION: chai>. 

as thorough admixture as possible of the culture material 
with the lymph emulsion, cover-glass specimens were 
made from the mixture, and stained in appropriate 
fashion. All of these preparations on subsequent ex- 
amination by the microscope showed large numbers of 
tubercle bacilli in every field. Both tubes were then 
set aside for a month, at the end of which the tubercle- 
containing vaccine lymph emulsion was employed for 
subcutaneous injection of guinea-pigs, other animals 
being at the same time injected with equivalent portions 
of the original growth of tubercle in order to serve as con- 
trols. These latter in due course developed tuberculosis 
locally, which subsequently became generalised, while, 
on the other hand, the animals which had been in- 
oculated with the tubercle -containing vaccine lymph 
emulsion showed no result either locally or generally. 
The question of quality of vaccine lymph naturally 
\ engaged the attention of the English Royal Commission 
on Vaccination at the time they were taking the 
evidence of witnesses, and among the recommendations 
contained in their final report is one to the effect that 
no persons should be required to submit to vaccination 
by means of any other lymph than that derived from 
the calf. The reason for this recommendation appears 
to be avoidance of even remote risk of inoculation, in 
the process of vaccination, of infections other than cow- 
pox — such, for instance, as those of syphilis and leprosy 
(sect. 433) — as also the desirability of minimising the 
opportunity for infection by erysipelas by obviating the 
necessity for opening vaccine vesicles involved in arm- 


to-arm vaccination (sect 447). The succeeding section 
of the report, which deals specially with the subject 
now being discussed, may be set out in full. It is as 
follows : — 

448. We think that safety would be increased by 
preserving the lymph in tubes instead of on 
" dry points." There is some difference of 
opinion on this matter among those with 
whose opinions we have been furnished. 
On the whole, however, we think the weight 
of experience as well as reason is in the 
direction we have indicated. 

In connection with this subject, our atten- 
tion has been drawn to the experiments 
recently made by Dr. Copeman as to the 
effect of the storage of vaccine lymph in 
glycerine. The conclusions at which he 
arrives are that the addition of glycerine, 
whilst it leaves the efficacy of the lymph un- 
diminished, or even increases it, tends to 
destroy other organisms. If it be the fact 
that the efficacy of the lymph remains unim- 
paired, its storage in glycerine would largely 
diminish the difficulties connected with the 
use of calf lymph which are inseparable from 
calf-to-arm vaccination. The investigation 
has not reached a point at which it is 
possible to pronounce with certainty whether 
the anticipated results would be obtained. 
And it was at one time suggested that the 

I70 VACCINATION : chap. 

introduction of glycerine was likely to be 
mischievous. The question is one a further 
investigation of which is obviously desirable. 
If lymph is to be preserved in glycerine, due 
care would be requisite to ensure its purity 
and the absence of contamination in its 
introduction. We think that, whether mixed 
with glycerine or not, each tube should 
contain only sufficient lymph for the vaccina- 
tion of one person. (August 1896.) 
In this section (448) of the Royal Commissioners' 
Report, a sentence occurs to which some special reference 
is desirable. This is as follows : — " And it was at one 
time suggested that the introduction of glycerine was 
likely to be mischievous." The reference made is to an 
outbreak of disease having the clinical characters of 
impetigo, which occurred in the summer of 1885, in 
villages situated on the island of Riigen, in the Baltic, 
after the vaccination of seventy-nine children in the 
latter part of June of that year. 

This outbreak forms the subject of a paper by the 
late Sir George Buchanan in the Transactions of the 
Epidemiological Society ioi 1885-86. He states that for 
the vaccination of these seventy-nine children only two 
tubes of lymph (of human origin) were available, and 
consequently their contents were further mixed with 
glycerine (glycerinum purissimum) before use. Thymol, 
to the extent of one-third, had also been mixed with 
the lymph previous to its having been sent out from the 
Government EstabUshment at Stettin. Sir George 


Buchanan states that the principal misgivings of the 
Commission appointed to inquire into the outbreak had 
concern with the glycerine, but adds that several points 
in the circumstantial evidence, however, suggest that the 
Stettin mixture, before glycerine had been added, may have 
been responsible for some irritation. He further says, 
however, " I do not know of any experience in England 
of inoculation with thymol or thymol and glycerine, but 
I have heard of dilutions of lymph with glycerine, always 
from people complaining of the lymph." 

In giving evidence before the Royal Commission on 
Vaccination in 1893, 1 was asked whether this statement 
did not condemn my suggested use of glycerine in 
connection with lymph supplies. I pointed out, how- 
ever, that, in the first place, dilution of lymph with an in- 
definite quantity of "glycerine" of unknown composition 
just before employing the mixture for vaccination, was a 
very different matter from storing lymph with chemically 
pure glycerine in definite amount and of definite 
strength, the whole being protected from light and air 
for a length of time before use, and that, in addition, 
what at the time of the outbreak in question may have 
been termed glycerinum purissimum, would in all 
probability at the present day be considered by no 
means deserving of that term. This latter statement I 
made on the authority of Messrs. Price, the chief 
manufacturers of glycerine in this country, whose 
manager, at the request of the directors, was good 
enough to put at my disposal his extensive practical 
knowledge of the details and the literature of this 

172 VACCINATION : chap. 

subject. The information thus obtained has been 
published in the appendix to the last volume of 
evidence, recently issued by the Royal Commission 
on Vaccination. 

The glycerine which has been employed in my in- 
vestigations is that manufactured in this country by 
Messrs. Price, but in Germany, where this method of 
purification and preservation is now carried out in all 
the Government Establishments for the supply of 
vaccine lymph, a glycerine made by Sarg of Vienna 
is used. This is said by Dr. Schultz, the Director of 
the Berlin Establishment, to be of a less " drying " 
nature than English glycerine. 

In order to obtain an authoritative opinion as to 
the degree of chemical purity obtained in the manu- 
facture of glycerine at the present time, and also 
an accurate estimate of the difference, if any, in the 
nature of the English and Austrian brands, I requested 
Dr. Wilson Hake to analyse samples of glycerine manu- 
factured by Messrs. Price and by Sarg respectively, and 
also samples of a new English brand which had just 
been placed on the market by Messrs. Lever Brothers. 
The result of the chemical examination of these 
samples, which Dr. Hake was kind enough to make for 
me, proves incontestably that all three samples exhibit a 
high degree of purity, most markedly so in the case of 
Messrs. Price's manufacture. In all there is complete 
absence of metallic contamination of any kind, while 
the amount of organic impurity is insignificant. The 
main difference observed is the slightly greater degree 


of concentration obtained in the English products as 
opposed to that of Austrian manufacture. 

In accordance with the suggestion contained in 
section 448 of the Report of the Royal Commission, 
the Local Government Board instructed me to make 
arrangements with Dr. Blaxall, the Lecturer on Bacteri- 
ology at Westminster Hospital, and now Bacteriologist 
to the Government Glycerinated Lymph Laboratories, 
to carry out a further series of experiments on the lines 
indicated by me in the evidence which had been tendered 
to the Commission, with the object of elucidating more 
fully the exact value of my glycerine method for the purifi- 
cation and preservation of vaccine lymph. The results 
of this series of experiments are set out in the Report 
of the Medical Officer to the Local Government Board 
for the year 1895-96, published in the autumn of 1897. 

Although devoting our attention mainly to the 
results which ensued on the employment of glycerine, it 
was thought desirable that investigation should be made 
concurrently as to the possible value for the preser- 
vation and purification of vaccine material, of lanoline 
and vaseline, substances which within quite recent years 
have been introduced by two officers of the Indian 
Medical Service, Surgeon Lieut. -Colonel King, and 
Surgeon-Major Bamber respectively, as being superior 
in their opinion to glycerine as agencies for the preser- 
vation of vaccine.^ Neither of these observers, how- 

^ The use of vaseline as an excipient for vaccine material had appar- 
ently been advocated by Professor Layet of Lyons, some years pre- 
vious to Surgeon-Major Bamber's first communication on the subject. 


ever, has as yet brought forward any bacteriological 
evidence as to a possible germicidal action of the 
substance they advocate, as will be seen on reference 
to their original papers. And Dr. Blaxall has shown 
that from this point of view, at any rate, neither lanoline 
nor vaseline can be regarded as possessing any value 
whatever. Indeed, it would appear that the extraneous 
organisms originally present in the lymph, with which 
these substances are admixed, so far from exhibiting as 
time goes on any diminution in numbers, tend, on the 
contrary, to become largely increased, as shown by the 
test of plate-cultivation. 

Lanoline and vaseline are thought by their advocates 
to possess superiority over glycerine mainly for the follow- 
ing reason. These officers note that glycerine emulsion 
tends to become mouldy after a short period, whereas 
this they do not find to be the case when either lanoline 
or vaseline is employed. That glycerinated lymph 
should have suffered deterioration in the way they indi- 
cate suggests either that it was not properly prepared, 
or that it was left exposed to the air. That the oc- 
currence of a similar accident is not an impossibility 
with lanoline and vaseline preparations, has been demon- 
strated by Dr. Blaxall's recent experience of their use. 

As examples of the results obtained, I set out in full 
the record of two experiments in which the action of 
glyceri?ie, vaseline, and lanoline respectively on vaccine 
material is compared. I append also the record of one 
experiment in which glycerine only was employed, but in 
which the glycerinated vaccine pulp was employed by 


Dr. Cory for the vaccination of a considerable number 
of children at the Government Vaccine Establishment, 
Lamb's Conduit Street. 

The glycerine used was Price's pure glycerine, 
having a specific gravity of 1200. It was thoroughly 
mixed with sterilised distilled water to 50 per cent, 
being weighed, and the resulting mixture was again 

The vaseUne used was pure vaseline, which, when 
bacteriologically tested, was found to be sterile. The 
lanoline used was pure anhydrous lanoline, and this 
also had been found to be sterile. 

The vaccine material was collected from calves which 
had been vaccinated in the ordinary course at the 
Animal Vaccine Establishment, Lamb's Conduit Street. 
Lymph from these same calves was being used as the 
current supply at that station. 

The vaccine material, which was collected 120 hours 
after vaccination of the calf, was procured in the 
following manner : the vaccinated surface of the calf 
was well cleansed with water, and the crusts on the 
vesicles wiped off with a clean towel. Next the vesicles 
were scraped with a sterilised spoon, and the scrapings 
and vesicular contents were received into a sterilised 
stoppered bottle of known weight. 

This bottle with its contents was then immediately 
taken to the laboratory and the experiment proceeded 

Experiment i. — The vaccine material (120 hour 
lymph) from the bottle was divided into three equal 


parts by weight, the division being effected expeditiously 
under cover and with sterilised instruments, so that no 
contamination of the vaccine might take place. Each 
separate portion of lymph was then rubbed up in a 
sterilised mortar (as before under cover) and was mixed 
with the chosen vehicle for its dilution. Complete 
distribution of the vaccine material throughout the 
vehicle was obtained in each instance by prolonged 

In this way : — 

One part was mixed with four times its weight of 

50 per cent glycerine in distilled water, so that 

the resulting emulsion contained 40 per cent of 

One part was mixed with four times its weight of 

vaseline undiluted, and the resulting emulsion 

contained 80 per cent vaseline. 
One part was mixed with five times its weight of 

lanoline undiluted, so that the resulting emulsion 

contained 83.3 per cent of lanoline. 
Immediately that satisfactory emulsions had been in 
each instance obtained, a definite amount of nutrient 
agar-agar, contained in a test-tube which had just pre- 
viously been liquefied and then allowed sufficiently to 
cool, was inoculated with emulsion, was well shaken up, 
and poured on a plate. Inoculation of the agar was 
effected by means of a looped platinum needle — the 
same needle with the same loop being used for all the 
experiments. The number of loopfuls taken from any 
one emulsion was intended to be equivalent to one 

9 i r 


Fig. 3. — Photograph of agar plate prepared with vaccine material 
immediately after admixture with Vaseline. 

Fig. 4. —Photograph of similar agar plate prepared with vaccine 
material six weeks after admixture with Vaseline. 


loopful of the undiluted vaccine material. Thus, of the 
diluted glycerine emulsion five loopfuls were taken, of the 
vaseline emulsion five, and of the lanoline emulsion six. 

Three plates of nutrient agar-agar were established 
in the above fashion from each emulsion, the second 
plate being seeded through the medium of three loop- 
fuls of material from the test-tube whence the first 
plate was poured, and the third plate being established 
in like manner from the test-tube for the second plate. 
The stock vaseline and lanoline emulsions were then 
placed in sterilised bottles duly stoppered. The stock 
diluted glycerine emulsion was placed in a sterilised test- 
tube plugged with cotton-wool. All were stored in a 
cool cupboard in the dark. 

The several agar plates were, after solidifying, placed 
for twenty-four hours in an incubator at 20° C, and then 
for six days in an incubator at 20° C. At the expira- 
tion of that time the several plates were photo- 
graphed. In this first experiment it was found, after 
six days' incubation at 20° C, that the first or original 
agar plates in all three series were so densely crowded 
with colonies as not to promise good pictures. Where- 
fore the second plates or first dilutions of the original 
agar tubes were alone photographed. A rough counting 
of the colonies on these plates, which had been estab- 
lished immediately after the emulsification had been 
effected, gave the following numbers : — 

Glycerine, Plate II. . . 1000 colonies per plate 

Vaseline, Plate II. . . 400 colonies per plate 

Lanoline, Plate II. . . 700 colonies per plate 


178 VA COIN A TION : chap. 

One week after the emulsificatiorl, further plates were 
established in the same manner, with the same number 
of loopfuls for thie . stock emulsions. Those plates 
showed an increase in the number of colonies in the 
samples from the vaselinated and lanolinated emulsions, 
but a decrease of colonies in the samples from the 
glycerinated emulsion. Similarly with plates made a 
fortnight and three. weeks after making the emulsions, 
the colonies from the glycerinated material were fewer, 
whilst those from the vaselinated and lanolinated 
sources were much more numerous than before. At the 
end of four weeks none of the plates established from the 
glycerine emulsion showed any colonies at all, while at 
this date plates from the vaseline and lanoline emulsions 
still showed increase of colonies. At the" end of six 
weeks plates from the glycerine stock emulsion were 
again found free from extraneous organisms, whereas 
the plates from lanoline and vaseline stock emulsions 
were crowded with colonies. A rough counting of 
these plates gave at this date : — 

Glycerine, Plate I.^ . . o colonies per plate 

Vaseline, Plate II. . 8,000 colonies per plate 

Lanoline, Plate II. . . 10,000 colonies per plate 

Experiment 2, — Scrapings of vesicles were obtained 
from a calf ninety-six hours after vaccination, the same 
precautions being taken as before. This material was 
weighed and divided into three equal parts. One part 
was emulsified with four times its weight of 50 per cent 
glycerine and distilled water, one part with four times 

1 Plate I. — This was so ; see next page. 



5. — Photograph of agar plate prepared with vaccine material 
immediately after admixture with Lanolink. 

Fig. 6. — Photograph of similar agar plate prepared with vaccine 
material six weeks after admixture with Lanoline. 


its weight of vaseline undiluted, and one part with five 
times its weight of lanoline undiluted as in the first 
experiment. Agar- agar plates were established from 
these emulsions antecedent to their storage, and the 
number of colonies which appeared on the several 
plates at this stage were found in close agreement. The 
three emulsions were then placed in sterile stoppered 
bottles, some of the glycerinated material being also stored 
in capillary tubes and pipettes. It was designed to repeat 
the former experiment with these emulsions, and sample 
plates were again established from them after they had 
been stored a week. Again the vaseline and lanoline 
plates showed at this stage increase in the number of 
colonies, whereas the glycerine plate showed a diminution. 
The vaseline plate, moreover, after four days, became 
covered with colonies of a green mould (Penicillium 
glaucum), and was useless for photographic purposes. 

At the end of the second week of this experiment, it 
was found impossible to continue it as regarded the 
vaseline and lanoline emulsions. These had become, 
in their separate steriHsed bottles, both covered with a 
luxuriant crop of the above green mould. No such 
growth appeared in the bottle containing the glycerine 
emulsion ; on the contrary, plates •established from it 
after two and three weeks' storage showed a further 
diminution in the number of colonies, and at the end 
of the fourth week the comparison plate was altogether 
free from extraneous organisms. 

In the foregoing experiments it will have been noted 
that the plates used for comparison were, in nearly 


every instance, plates established from agar test-tubes 
directly inoculated from the emulsion. The exception 
to this rule was Experiment i, in which the plates used 
for comparison were established from agar tubes inocu- 
lated, not directly from the emulsions, but from the 
primary agar tubes in which loopfuls of emulsion had 
been, in the first instance, distributed. In that experi- 
ment the sterile plate obtained at the end of the fourth 
week of storage of vaccine material in glycenne was the 
only plate of the series established directly from the 
original emulsion which has been photographed. 

It remains to be stated in this connection that the 
results were " controlled " by duplicate plates established 
at the same time ; and furthermore, that material which 
in plate culture showed no colonies was always tested 
again two or three times in similar fashion. 

Experifnent 3. — Scrapings of vesicles were collected 
from a calf 120 hours after vaccination, with the same 
precautions as before. These were weighed, rubbed 
up, and diluted with fifteen times their weight of 50 per 
cent sterile glycerine and water, so that the percentage 
of glycerine present became 46.9. One loopful of this 
emulsion immediately after glycerination was inoculated 
into nutrient agar -agar, and plates were established 
therewith. Plates established from the stock glycerine 
emulsion a week, a fortnight, three weeks, and four 
weeks respectively from commencement of storage, 
showed, as regards the first three, week by week 
a rapid diminution in the number of colonies which 
appeared after incubation in the manner described. 


while the fourth was found to be quite free from 
extraneous organisms. Some of this glycerine emulsion, 
four weeks after manufacture and storage in a pipette, 
was used by the Government operators at the Animal 
Vaccine Establishment, Lamb's Conduit Street, for the 
vaccination of sixty -five infants. All these children's 
arms "took," Dr. Cory obtaining in his series of thirty- 
six cases loo per cent of insertion success. 

The advantages of glycerinated calf lymph may be 
set out as follows : — 

1. By employing the method of glycerination of 
lymph-pulp, great increase in quantity can be obtained 
without any consequent deterioration in quality, the per- 
centage of insertion success following on its use being 
equal to that obtained with perfectly active fresh lymph. 

2. Glycerinated lymph does not dry up rapidly, as 
does unglycerinated lymph, thus simplifying the process 
of vaccination. 

3. Glycerinated lymph does not coagulate, so that it 
never becomes necessary to discard a tube on this account. 

4. Glycerinated lymph can be produced absolutely 
free from the various streptococci and staphylococci 
which are usually to be found in untreated calf lymph, 
and which are, under certain circumstances, liable to 
occasion suppuration. 

5. In like manner the streptococcus of erysipelas, 
in the event of its having been originally present in the 
lymph material, is rapidly killed out by the germicidal 
action of the glycerine. 

6. The tubercle bacillus is effectually destroyed even 


when large quantities of virulent cultures have been 
purposely added to the lymph. 

7. The possibility of inoculation of syphilis is elim- 
inated, as the calf is not subject to this disease. 

8. The necessity for collecting children together, 
with the attendant risk of spread of infectious diseases, 
or of transporting a calf from place to place, is obviated, 
while the danger of "late" erysipelas in the child is dim- 
inished by reason of there being no necessity to open 
the mature vesicles for the purpose of obtaining lymph. 

9. The bacteriological purity and clinical activity of 
large quantities of the lymph can be readily tested prior 
to distribution. 

10. By reason of the possibility of keeping large 
stocks of glycerinated lymph on hand for considerable 
periods of time without appreciable deterioration, any 
sudden demand, such as is likely to arise on the outbreak 
of epidemic smallpox, can be promptly met. 

11. The expense of producing glycerinated lymph 
is proportionally small, since the amount obtainable 
from each calf is enormously increased. 

The Preparation of Glycerinated Calf Lymph 

The method best adapted for the production of 
glycerinated calf lymph, which shall be free from 
all extraneous organisms, of perfect efficacy, and yet 
affording material for the vaccination of many more 
children than the original unglycerinated calf lymph, 
is briefly as follows : — 

The Preparation of the Calf. — A female calf of suit- 


able age, about from three to six months, should be 
kept under observation for a week, after which, if found 
to be quite healthy, it may be removed to the vaccina- 
tion station. It is there placed on a tilting table, and 
the lower part of the abdomen, reaching as far forward 
as the umbilicus, is shaved and thoroughly washed with 
a solution of carboHc acid and then rinsed with sterile 
water and dried with soft sterilised towels. 

Inoculation of the Calf. — With a sterilised sharp scalpel 
incisions about four inches long and half an inch apart, 
parallel to the long axis of the body, are made on this 
clean-shaved area. The depth of the incision should 
be such as to pass through the epidermis and to open 
the rete Malpighi, if possible without drawing blood. 
As these incisions are made, glycerinated calf lymph, 
which by examination has been proved to be free from 
extraneous organisms, is run into them by means of a 
sterilised blunt instrument, and the point of the scalpel 
is from time to time dipped into the vaccine emulsion. 

Collection from the Calf. — After five days (120 hours) 
the vaccinated surface of the calf is first thoroughly 
washed with warm water and soap, rubbed over it by 
the clean hand of the operator, and finally the whole 
area is carefully cleansed with sterile water. The 
remaining moisture is then removed by steriHsed sheets 
of blotting paper. The vaccinated incisions will now 
appear as lines of continuous vesicles raised above the 
surface, each line separated from its neighbour by about 
a quarter of an inch of clear skin. Any crusts which 
appear in the vesicular lines are picked off with a blunt 


sterilised instrument. The vesicles and their contents 
are then removed by means of a sterilised Volkmann's 
spoon, and transferred to a sterilised bottle of known 
weight. By going over the lines only once with the spoon, 
it is quite easy to remove the whole of the pulp without 
any admixture of blood. The abraded surface is care- 
fully washed, and may be dusted over with fine oatmeal 
or starch and boracic powder. Subsequently, the calf 
is transferred to the slaughter house and the carcase is 
examined by the veterinary surgeon, who forwards a certi- 
ficate of its condition. Should this not be satisfactory, 
the vaccine pulp obtained from the animal is destroyed. 
Preparation and Glycerinatio?i of the Lyniph-pulp. — The 
bottle containing the vaccine pulp is taken to the 
laboratory, and the exact weight of the material ascer- 
tained. A calf vaccinated in this way will yield from 
18 to 24 grammes, or even more, of lymph-pulp. This 
material is then thoroughly rubbed up in a sterilised 
mortar or in a mechanical triturating machine.^ When 
it has been brought to a fine state of division, it is mixed 
with six times its weight of a sterilised solution of 50 per 
cent chemically pure glycerine in distilled w^ater. The 
resulting emulsion is then transferred to small test-tubes, 
which are then aseptically sealed and should be stored in 
a cool place protected from light. When required for 
distribution it is drawn up into sterilised capillary tubes, 
which are subsequently sealed in the flame of a spirit lamp. 

^ For description of instruments devised by Dr. Chalybaus of 
Dresden, and Dr. Doering of Berlin, respectively, see Appendix I. 
pp. 222 and 213. 


Series 24. — Photograph of agar plate incubated for seven days sub- 
sequent to inoculation, with one loopful of rubbed-up vaccine pulp, im- 
mediately after glycerination. The number of colonies is very large. 


Series 24. — Photograph of agar plate prepared with similar quantity of 
vaccine material one week after glycerination. The number of colonies is 
considerably diminished. 


Series 24. — Photograph of similar agar plate, prepared with vaccine 
material two weeks after glycerination. A further decrease in the number 
of colonies is now conspicuous. 


Series 24. — Photograph of similar agar plate, prepared with vaccine 
material three weeks after glycerination. The decrease in the number of 
colonies is still more marked than in the former plates. 


Series 24. — Photograph of similar agar plate, prepared with vaccine 
msLterial /our weeks after glycerination. No growth whatever has occurred.. 


Series 24. — Photograph of similar agar plate, prepared with vaccine 
maXenaX four weeks after glycerination. No growth whatever has occurred.. 


Bacteriological Examination of the Lymph Emulsion. — 
As soon as the vesicular pulp is thoroughly emulsified 
with the glycerine solution, agar-agar plates are estab- 
lished from it, and, after suitable incubation for seven 
days, the colonies that have developed on the plates are 
counted and examined. Week by week this process 
is repeated, and invariably the number of colonies 
diminishes with the age of the emulsion, until at the 
end of the fourth week after the collection and glycerin- 
ation of the lymph material, the agar-agar plates inocu- 
lated at that time show no development of colonies. 
The lymph is then subjected to further culture experi- 
ments, and if these results of freedom from extraneous 
organisms are confirmed, the emulsion is ready for 
distribution. The elimination of the extraneous organ- 
isms in our experiments has occurred with marked 
regularity at the end of the fourth week. The only 
exception to this rule arises when the lymph originally 
contained a considerable number of spores or bacilli of 
the hay bacillus or bacillus mesentericus. These organ- 
isms are very resistant to the action of glycerine, but if 
the precautions detailed are carried out in the treatment 
of the calf, their presence may generally be excluded. 

Duration of Activity of Glycerinated Calf Lymph. — 
This varies in all probability with atmospheric condi- 
tions, with the fineness of division of the vesicle pulp, and 
above all with the condition of the calf itself Some 
calves yield an excellent lymph, others a poor lymph, 
and the problem is to determine the value of the lymph 
yielded by any given calf A lymph which was collected 

1 86 VA CCINA TION : chap. 

and glycerinated on 13th July 1897, has since been used 
at intervals of from twenty-four weeks to thirty-two weeks 
after glycerination, for the vaccination of children. Dur- 
ing this period, sixty-one children have been vaccinated 
with this lymph in five places each, with a mean inser- 
tion success of 98 per cent.^ Thus, by the methods 
described, glycerinated calf lymph can be prepared which 
becomes freed from extraneous organisms, is available 
for a large number of vaccinations, at least 5000 from an 
average calf, and retains full activity for eight months, 
and will, under favourable circumstances, continue to do 
so in all probability for still longer periods, if necessary. 

From the description which I have given, it will, I 
think, be obvious that the preparation and testing of 
glycerinated calf lymph, properly so called, is a matter 
requiring considerable skill and care. Sir Richard 
Thorne insists strongly on this fact in his introduction 
to our joint Report, recently presented to Government, 
on the administration of certain of the chief continental 
vaccine estabUshments. This Report I reproduce as 
an appendix, with such -emendations and additions as 
more recent information and further experience have 
shown to be needed. 

The following conclusioris were submitted by Sir 
Richard Thorne in his introduction to the Report in 
question : — 

" I. It is desirable that vaccination, both primary and 

^ This same sample of lymph (when forty-two weeks old) gave, 
in Dr. Cory's hands, an insertion success, based on fifty insertions, 
of 100 per cent. 


Photograph of child, taken on the eighth day of vaccination with glycer- 
inated vaccine lymph, which, as shown in Plate I. Fig. 2, was found, when 
tested by the method of plate -cultivation, to be free from extraneous 


secondary, carried out under the auspices of the Govern- 
ment, should be performed exclusively with vaccine 
lymph derived from the calf. 

"2. There will probably be advantage in retaining, 
for a time at least, the system of calf-to-arm vaccina- 
tion at the Board's Animal Vaccine Station for such 
parents and others as may request it, and for the pur- 
poses of comparing its results with those following the 
use of calf lymph preserved in one or another way. 

"3. The distribution of calf vaccine from the National 
Vaccine Establishment should be limited to glycerinated 
or similar preparations of lymph or pulp material, in 
air-tight tubes or other glass receptacles. 

"4. To give effect to the above, it will be requisite 
that the Board's Animal Vaccine Station should be 
reorganised, both as regards construction and adminis- 
tration. Notably will it be requisite that it should 
include a properly equipped laboratory under the direct 
supervision of a bacteriological expert." 

[These recommendations are now in process of 
adoption by the Government, and already suitable 
laboratories have been secured, which have been placed 
in the charge of Dr. Blaxall. I trust, therefore, that 
the Government will shortly be in a position to supply 
bacteriologically pure and fully active calf vaccine lymph 
in any quantity that may be found desirable.] 


Preservation, Storage, and Use of Lymph : i. Ad ami. 

Montreal Medical Joicrn., Jan. 1898, vol. xxvii. p. 52. — 2. 
Blaxall. Medical Magazine, April 1898, vol. vii. p. 253. — 3. 
BOUSQUET. Nouveau traits de la vaccine, p. 240. — 4. Chambon, 
Menard, and Straus. Gazette des hdpitaux, Dec. 15, 1892. — 
5. Cheyne. Medical Times, Old Series, 1850, vol. xxi. ; and 
New Series, 1853, vol. vi. — 6. Copeman. Trans, of Inter- 
national Congress of Hygiene, 1 89 1, vol. ii. p. 325 ; Proceedings 
of Royal Society, 1 893, vol. liv. p. 187; Brit. Med. Journ. 1893, 
vol. i. p. 1250. — 7. Copeman and Blaxall. Report of Med. 
Officer to Local Government Board, 1895-96, pp. 283-305. — 

8. Dreyer. Zeitschrift fiir Hygiene, 1898, Bd. xxvii. p. 117. — 

9. Leonl Revue d'hygiene, Aug. 20, 1894. — 10. Muller. 
Med. Times and Gazette, 1866, May 19, p. 526. — ii. Seaton. 
Handbook of Vaccination, 1868 ; Second Report of Med. Officer to 
the Privy Council, p. 20 et seq. — 12. Steinbrenner. Traitt^ 
sur la Vaccine, p. 570. — 13. Warlomont. Brit. Med. Journ. 
1880, vol. ii. p. 499. — 14. Second Report of Med. Off[icer to Privy 
Comtcil. — 15. Fifth Report of Med. Officer to Privy Council. 


On the Preparation and Storage of Glycerinated 
Calf Lymph : by Drs. Thorne Thorne and 
Monckton Copeman 

Introduction addressed by the Medical Officer to the Right 
Honourable Henry Chaplin, M.P., President of the 
Local Government Board 

Sir — Shortly after the issue of the Report of the Royal 
Commission on Vaccination last autumn, I received your 
instructions that I should, together with Dr. Monckton 
Copeman, visit certain cities in different countries on the 
Continent of Europe, with a view of obtaining information 
as to the methods adopted, by the respective authorities and 
others concerned, in the distribution of vaccine lymph derived 
from the calf, more especially in reference to the prepara- 
tion, storage, and distribution of glycerinated calf lymph. 

The necessary visits were commenced in December of 
last year, but owing to the fact that in some of the countries 
to be visited public vaccination is practically limited to 
certain months of the year, commencing with the spring, it 
was found impossible to complete the inspections until quite 
recently. In eliciting information as to the methods adopted 
in each of the countries visited, we held especially in view 
two points to which the Royal Commission gave prominence 


in their Report. The first of these — one which the Com- 
missioners put " in the forefront," is the recommendation in 
section 437 of their Report "that parents should not be 
required to submit their children to vaccination by means 
of any but calf lymph." The second, which is referred to 
in section 448, is concerned with experiments which Dr. 
Copeman made and announced to the International Congress 
of Hygiene which met in London in August 1 891, as to the 
effect of the storage of vaccine lymph in glycerine. " The 
conclusions at which he arrives," say the Commissioners, 
"are that the addition of glycerine, whilst it leaves the 
efficacy of the lymph undiminished, or even increases it, 
tends to destroy other organisms " ; and they add that 
" The question is one a further investigation of which is 
obviously desirable." 

The places visited by us were Paris, Brussels, Berhn, 
Dresden, Cologne, and Geneva ; and in submitting to you 
an account of the operations which came under our notice 
in each of these cities, I beg leave to make the following 

It will be noted that in each of the countries concerned, 
vaccination with calf lymph has become the habitual, if not 
the universal practice. In some, indeed, we were informed 
that, although vaccination with humanised lymph is not 
definitely prohibited under any statute or regulation, yet 
resort to such lymph by any medical practitioner having 
official responsibility to the Government is altogether 

In only one of the places visited — namely, Paris — did 
we find that vaccination was carried out under official 
sanction with crude calf lymph, and even there the process 
was limited to vaccination direct from calf to arm, all lymph 
stored for distribution being glycerinated calf lymph. 


The circumstances of Paris in the matter of vaccination 
direct from the calf deserve consideration, because they 
have a certain similarity to those which may be encountered 
in this country, in case the use of calf lymph should become 
habitual. Thus, we were informed that if vaccination had, 
during past years, been limited to the use of glycerinated 
calf lymph, stored in tubes or otherwise, it would have been 
found difficult, amongst certain classes of a population which 
heretofore had only known of stored humanised lymph, to 
convince persons whose vaccination it was desirable to 
secure that the lymph proposed to be used on them was 
really calf lymph, not humanised lymph. In order to 
ensure confidence in this respect, it had been the practice 
to convey calves to the vaccination stations, or to districts 
infected with smallpox, and to perform the vaccination direct 
from calf to arm. Perusal of the account which we give as 
to this practice in the hands of MM. Chambon and Menard 
will show that it has been brought to a position of very 
considerable efficiency. 

In all the other countries visited we found that, acting 
on the indications announced by Dr. Copeman in 1891, 
the Governments and other authorities concerned had made 
sustained investigation as to the preparation, storage, and 
use of glycerinated calf lymph, and had gradually come to 
adopt that preparation of lymph for official, and in some 
cases for all, purposes throughout their jurisdictions. Thus, 
in Germany we were informed that the system of vaccinating 
direct from the calf had come to be abandoned as completely 
as that from arm to arm, the use of glycerinated calf lymph 
having become general throughout the Empire. 

The reasons for this change have been two. The 
governing reason has been the confirmation by competent 
bacteriologists of the results obtained by Dr. Copeman, to 


the effect that, by the admixture to calf lymph of a 50 per 
cent solution of pure glycerine in sterile water, and by 
subsequent storage of the lymph material in tubes, under 
due precautions, for a term of several weeks, the preparation 
remained quite active as vaccine, whilst a very remarkable 
germicidal effect was produced on extraneous micro- 
organisms in the lymph, even including certain pathogenic 
organisms which had been purposely added to the lymph 
material. The second reason was that, by reason of the 
admixture referred to, the amount of vaccine procurable 
from a given calf could be greatly, even enormously, in- 
creased, and that, within certain wide limits, this could be 
done without interfering with the insertion success following 
on the use of the lymph. At the Board's Animal Vaccine 
Establishment it has hitherto not been deemed necessary, 
nor even expedient, to make one calf serve for more than 
some 200 to 300 vaccinations. It is no unusual thing abroad 
to provide from a single calf an amount of glycerinated 
lymph that shall serve for from 4000 to 6000 vaccinations, 
and in Berlin we were assured that the glycerinated lymph 
which was prepared in our presence from one calf would 
suffice for no less than 15,000 vaccinations. We brought 
some of this Berlin lymph to England, and it was used for 
the purposes of vaccination at intervals of nine, eleven, 
and thirty-seven days after its collection, with the result 
that in 76 vaccinations performed, in each case by five 
insertions, its use resulted in a mean insertion success 
of 92.0 per cent. Storage of this particular sample for a 
much longer period did not give satisfactory results. With 
other preparations of glycerinated lymph, diluted to about 
one-third of the amount of the Berlin sample, and which 
were used in England at intervals varying from seven 
to thirty-one days after collection and preparation, the 


insertion success reached 97, 98, and 99 per cent ; and 
in the case of 1 1 1 vaccinations, all effected with two other 
supplies of glycerinated lymph, used at intervals of from 
seven to thirty days after preparation, the success reached 
100 per cent, every insertion of lymph being followed with 
success. In all these cases the vaccination was performed 
by means of five insertions. 

In every instance we found that the work of collecting, 
preparing, and storing the glycerinated lymph was carried 
out with the greatest care ; a condition of scientific cleanliness 
was especially aimed at, and a laboratory, fitted with bacterio- 
logical and other scientific apparatus, always formed an 
essential part of the vaccine institution. The extent to 
which the desired end of freedom from extraneous impurity 
was attained depended largely on whether a first attempt to 
adapt an existing calf station or similar establishment to its 
new purposes had been maintained, or whether it had been 
abandoned in favour of an institution constructed especially 
for the purposes of that which is in the main scientific 
laboratory work. Several of the stations which we visited 
are already under condemnation, because of the difficulty 
of ensuring that freedom from extraneous micro-organisms 
which should be aimed at during the preparation of the 
lymph supply ; the Cologne station is one of the newest, 
and may well serve, in its main features, as a type of that 
which should be aimed at. 

The condition of scientific cleanliness to which I have 
referred extended to such matters as the following : {a) the 
construction and administration of the stabling for the 
calves ; {b) the means for washing or bathing calves before 
their vaccination ; {c) the construction, cleansing, etc., of 
the operating rooms ; {d) the cleansing of the vaccinated 
surface of the calf with germicidal preparations and sterilised 



cloths before collection of the lymph ; {e) the use of clean 
sterilised outer garments by all officials concerned in the 
processes carried out ; (/) the sterilisation of all instruments, 
etc., employed ; and {g) the carrying out of the process of 
admixture of the lymph material or pulp with glycerine, as 
also its preparation and storage under conditions of laboratory 
freedom from extraneous organisms. 

Further, we found that it was an invariable practice at 
the stations visited on the Continent not to issue any lymph 
until a report had been received from a veterinary surgeon, 
after slaughter of the animal, as to the freedom of the calf 
furnishing it from disease ; in brief, the lymph issued is 
that of healthy calves only. This practice is mainly with a 
view of avoiding all risk of conveying tuberculosis along 
with calf lymph ; though such risk would, under any cir- 
cumstances, be a very remote one, seeing that tuberculosis 
is extremely rare in young bovine animals, and seeing also 
that the tubercle bacillus, when experimentally added to a 
mixture of lymph and an aqueous solution of glycerine, 
rapidly loses its vitality. 

The information which we obtained in the course of our 
visits does not profess to be complete. Much remains to 
be ascertained by careful scientific research, in order to 
learn what are the precise conditions under which glycerin- 
ated calf lymph can be prepared and stored, so as to secure 
to the utmost freedom from extraneous, and especially from 
pathogenic micro-organisms, whilst at the same time retaining 
to the utmost the undiminished protective value of the 
lymph material against smallpox. We learned that in every 
country visited, further research is being made in this 
direction, and in Germany a special commission of medical 
and bacteriological experts has been appointed by the 
Government to study and report upon the subject. 


But the information which is now available in this 
country, and that which, during the course of our visits 
abroad, was placed at our disposal with a readiness and a 
courtesy which calls for an expression of the fullest 
acknowledgment, suffices to enable me to submit the 
following conclusions for your consideration : — 

1. It is desirable that vaccination, both primary and 
secondary, carried out under the auspices of the Govern- 
ment, should be performed exclusively with vaccine lymph 
derived from the calf. 

2. There will probably be advantage in retaining, for 
a time at least, the system of calf-to-arm vaccination at the 
Board's Animal Vaccine Station for such parents and others 
as may specially desire it, and for the purposes of comparing 
its results with those following the use of calf lymph pre- 
served in one or another way. 

3. The distribution of calf vaccine from the National 
Vaccine Establishment should be limited to glycerinated or 
similar preparations of lymph and pulp material, in air-tight 
tubes, or other glass receptacles. 

4. To give effect to the above it will be requisite that 
the Board's Animal Vaccine Station should be reorganised, 
both as regards construction and administration. Notably 
will it be requisite that it should include a properly equipped 
laboratory, under the direct supervision of a bacteriological 
expert. — I have the honour to be, sir, your obedient servant, 


July 1897. 


APPENDIX No. \— Continued 

Report on the Results of an Inspection made by 
Dr. R. Thorne Thorne and Dr. S. Monckton 
copeman as to vaccination arrangements 


special reference to the preparation and 
Storage of Glycerinated Calf Vaccine 
Lymph. (Prepared by Dr. S. Monckton 


At Paris we spent several days in inspecting the manner 
in which the work of obtaining, preparing, storing, and 
distributing calf vaccine lymph is carried out at the Institut 
de Vaccine Animale and at the Academie de Medecine 

The Institut de Vaccine Animale 

This establishment is carried on by M. Chambon and 
Dr. St. Yves Menard. It is, practically, a private estab- 
lishment, although the municipality of the city of Paris 
contract with the directors to carry out all such public 
vaccinations, within their jurisdiction, as may be necessary. 

The Institut de Vaccine Animale, which is situate in 
the Rue Ballu, consists of what was originally a dwelling- 
house, with a courtyard opening to the street alongside, and 
a stable behind, the portion of the courtyard immediately 
adjoining the stable being covered over and provided with 
sliding doors, so as to form an operating room when neces- 


sary. Rooms in the dwelling-house on the ground floor are 
set aside as waiting and operating rooms in which persons 
are vaccinated direct from the calf, while on the first floor 
are other rooms in which the calf lymph is manipulated, 
placed in sealed tubes, stored, and distributed. 

Stable. — This is a building about 18 feet square, 
which contains stalls for ten calves. Each stall is some- 
what narrow, but we were informed that this was advan- 
tageous, as the animal was thus kept more quiet than would 
otherwise be the case. Attached to the halter of each 
animal is a large iron ring, which runs on a vertical iron 
rod let into the wall of the building, above and below. 
This arrangement was devised in order to prevent the 
calves from being able to lick the inoculated area of their 
body, while, at the same time, it does not prevent them 
from lying down. The floor of the stable, which is formed 
of roughened bricks, slopes slightly to a shallow drain on 
either side of a footway between the two rows of stalls. 
The building is heated artificially by means of hot-water 
pipes, and its ventilation is aided by means of an extraction 
shaft containing a lighted gas jet. At the time of our visit 
the temperature was about 1 5 ° C. The walls are covered 
with glazed tiles, and the floor is laid with bricks which are 
impervious to moisture. Both walls and floor are occa- 
sionally washed down by the aid of a spray of a solution of 
perchloride of mercury. 

Calves. — These animals, which appear to be in every 
respect well suited for the purpose of lymph propagation, 
are of a special breed, and are obtained from the Depart- 
ment of Correze in the southern half of France. They are 
all of a uniform reddish-brown colour, with fairly long, soft 
hair. Their skin, when exposed by shaving, is seen to be 
particularly smooth and supple, and it varies in colour from 


pink to a pale shade of brown. For the purpose of keeping 

free from urine the straw -bed which comes into contact 

with the surface of the body operated on, only cow-calves 

are employed, which vary in age from four to six months, 

the average being about eighteen weeks. Prior to being 

brought to the Institut stable, they are kept for about twelve 

days in a quarantine shed in the outskirts of Montmartre. 

They are all weaned at the age of two months, and they 

receive no milk or eggs while at the quarantine shed 

or at the Institut stable, the food of each calf consisting 


One litre of crushed oats, 

Two litres of bran, and 

Three kilos of hay 

twice daily, at about 6 A.M. and 4 p.m. At both places 
water is supplied from the town mains. 

The calves cost, on an average, about 125 francs, with 
an additional 22 francs for travelling, etc., making 147 
francs in all. After use, they are sold to a butcher, at a 
loss of from 30 to 40 francs. 

On the day after arrival at the stable the calves are 
vaccinated. Injection of tuberculin is not employed, as the 
directors consider this unnecessary, in view of the fact that 
immediately after collection of the lymph each calf is sent 
to the abattoir and slaughtered under the personal supervi- 
sion of a medical man, whose report is awaited before any 
of the lymph is distributed for use. In the event of any 
signs of tuberculosis being found, the whole of the lymph 
derived from this particular animal would be destroyed. 

Vaccination of Calves. — For this purpose one or more 
large tubes of glycerinated lymph, which have been kept 
for at least a month subsequent to its collection and storage, 
are employed, the directors considering that better results 


are thus obtained than if the operation were carried out 
directly from calf to calf. 

For the purposes of the operation the calf is strapped to 
a tilting table somewhat similar in design to those employed 
in this country, but the right hind leg of the animal is not 
elevated, and in consequence the mammary region is not 
exposed, indeed it is not utilised for inoculation. 

To prepare a surface for the insertion of the lymph the 
right side of the animal is thoroughly scrubbed with soap 
and hot water, and then shaved over an area extending 
between the internal edges of the fore and hind limbs, and 
from some 4 or 5 inches below the spinal ridge to the 
umbilicus. The shaved area is next washed with soap and 
hot water, then with a hot solution of boracic acid, and, 
finally, with plain hot water. It is afterwards dried with 
clean soft cloths. 

A number of superficial incisions, each about i inch 
long, are then made in a direction at right angles to the 
long axis of the body, and about a couple of inches one 
from another. The incisions of the several rows are made 
en echelo7i. The lancet employed for the purpose has a 
spear-headed blade, this shape being specially recommended 
by M. Chambon. Over each incision a drop of glycerinated 
lymph is allowed to fall from a glass tube, and the drop is 
rubbed in with the flat portion of the blade of the lancet. 
The process is carried out by one of the laboratory servants, 
and is a somewhat lengthy one. 

When the lymph has dried, the calf is removed from the 
table and taken back to its stall. 

Collection of Lymph. — The vaccine material is always 
collected on the sixth day. The calf is once more placed 
on the table ; or, if material is required for immediate use 
only, it is usually allowed to stand. The vaccinated area 


is washed with warm water and dried with clean soft cloths. 
Each vesicle is now clamped separately, and the crust first 
removed with a lancet, which is then wiped on a cloth 
pinned to the front of the clean cotton blouse which the 
operator has previously donned. 

The vesicle is then thoroughly scraped with the edge of 
a somewhat blunt lancet, and the resulting mixture of lymph, 
epithelial tissue, and blood is transferred to a small nickel 
crucible set in a wide wooden stand on a table close to the 
operator. The crucible is provided with a cover which is 
kept over it except at the moment when a further addition 
is made to its contents. The collection of all the vesicular 
material obtainable from one calf appears to take about 
three-quarters of an hour. 

To the pultaceous mass contained in the crucible there 
is added about an equal quantity of glycerine, which was 
described to us as " doubly rectified," but which appeared 
to be of very thin consistence as compared with the best 
English glycerine. No accurate measurement of the 
quantity employed is made. 

The mixture of pulp and glycerine is triturated in a 
mixing machine devised by Dr. Chalybiius of Dresden, the 
particular one that we saw being driven by a small electric 
motor. (A description of this machine by Dr. Chalybaus 
himself will be found on pp. 222-224.) 

The mixture, having thus been rendered thin and homo- 
geneous, is received in a clean steriHsed nickel crucible 
placed beneath the machine, but with a view of still further 
improving its appearance and of removing any extraneous 
matters, such as hairs, it is afterwards pressed through a 
small brass-wire sieve, consisting of extremely fine gauze, 
into an agate mortar. This is done by means of a bone 
spoon, and there is left on the surface of the gauze nothing 


but a very small quantity of epithelial tissue, together with 
a few hairs. The mixture is further triturated in the 
mortar with an agate pestle, and is then ready for filling 
into the tubes in which it is distributed. 

Storage of Ly^nph. — The lymph material is next drawn 
up into a sterilised glass syringe provided with a metal 
nozzle of such a size that the tubes to be filled can easily be 
fitted over it. Slight pressure on the piston of the syringe 
causes sufficient lymph to enter the tube. Each tube is 
filled about two-thirds full, and is then placed on a porcelain 
tray, pending its being sealed. This is done without delay 
by means of a blow-pipe, the air blast of which is provided 
by means of a pressure apparatus in one corner of the room. 
The tube is first tilted until the column of lymph occupies 
the .central portion, and it is then held in a horizontal posi- 
tion, while each end is successively placed in the blow-pipe 
flame, and, when sufficiently melted, drawn out by means 
of a pair of forceps, and so sealed. 

Distribution of Lymph. — The tubes when sealed are 
placed with a small surrounding of cotton-wool in small 
light metal tubes provided with a tightly-fitting cover. 
These cases, if sent out singly, are fitted into a block of 
wood grooved on one side, being kept in position by a 
paper label, which is gummed round the block, and which 
has on one side space for postal address and stamp, and, on 
the other, directions for use. 

A register is- kept of persons to whom supplies of lymph 
are sent, and of the calf from which each supply is derived. 

Academie de Medecine 

We also visited the vaccine station of the Academie de 
Medecine, of. which Dr. Hervieux is the director. The 
general principles on which this institution is conducted are 


so similar to those which have been set out at length in the 
description of MM. Chambon and Menard's establishment, 
that it is unnecessary to enter into similar details again. It 
should be mentioned, however, that at the Academie de 
Medecine the calves employed were not of the same breed 
as those used at the Institut Vaccinal, neither did the 
vesicles which we had an opportunity of seeing, on a single 
visit, appear to be quite equal in character to those which 
we observed at the latter institution. 

Calf-to-Arm Vaccination at Nanterre 

While in Paris we were afforded an opportunity of seeing 
an extensive series of vaccinations and revaccinations, carried 
out directly from calf to arm, at the Nanterre House of 
Detention. The calf, which arrived at the establishment in 
a closed van, was brought from the vaccine institute of 
MM. Chambon and Menard, and the vaccinations were 
carried out by M. Chambon and by one of the medical 
officers to the establishment. 

The operations were performed in a small square room, 
having a door at each of two opposite angles ; opposite the 
door of entry the calf was tied up to a post, and, in front of 
and facing it, an assistant took up his position in order to 
collect the lymph, by scraping slightly one of the vesicles on 
the calf s abdomen, to which compression forceps had been 
applied. On his right hand were placed a tray containing 
lancets and compression forceps, a rack for holding charged 
lancets, and a glass bowl containing a pad of cotton-wool 
floating in perchloride of mercury solution. On either side 
of him, and so arranged as to be able with ease to reach 
the tray, sat one of the two operators, each of whom had 
placed in front of him a chair for the vaccinee. 


The male inmates, each with his shirt sleeve on the left 
arm rolled up to the shoulder, were admitted by the door 
opposite the calf, an assistant, wearing an ambulance 
badge, giving the upper part of each person's arm a brisk 
rubbing with a cloth soaked in boracic acid solution as he 
entered, the arm being afterwards dried with another cloth. 
Two or three warders were also in attendance, who so 
directed the stream of inmates that each operation chair 
was refilled as soon as vacated. 

On an inmate seating himself, the operator took a 
charged lancet from the rack, with which he made three 
punctures in an oblique direction, just beneath the skin of 
the upper arm. The lancet was then dropped into the 
glass bowl containing the disinfectant, from which, in turn, 
it was removed by the assistant, who wiped it on a pre- 
viously sterilised cloth, and the instrument was then 

One assistant was thus able to keep the two operators 
supplied with charged lancets as rapidly as they were 
required, and it will be obvious that the whole scheme of 
operation had been well devised, and was skilfully and 
expeditiously carried out, when we mention that during our 
visit no less than 480 vaccinations were performed in the 
short space of thirty-nine minutes. Most of the vaccina- 
tions were revaccinations, some of the inmates having been 
submitted to the same operation on the occasion of previous 
admissions to this or similar institutions. 

Arrangements for Gratuitous Domiciliary 
Vaccination in the City of Paris 

As already stated, MM. Chambon and Menard are 
entrusted, by the municipality of Paris, with the carrying 


out of arrangements for the vaccination and revaccination 
of the inhabitants ; and on receiving information of the 
occurrence of smallpox in any part of the city, they make 
domiciliary visits for the purpose of offering vaccination to 
persons who may be unable or unwilling to attend the 
public stations. 

The notifications as to the existence of smallpox are at 
once sent on to MM. Chambon and Menard, who then 
make arrangements to attend at the house or neighbour- 
hood in question on the following day ; but in the meantime 
the day and time of attendance are intimated to the 
inhabitants by means of printed cards having blank spaces 
for the insertion of the necessary particulars. In Paris 
each " house," in most of the quarters occupied by the poor 
and the working classes, is made up of a series of flats, 
which, again, are subdivided up into dwellings of one or 
more rooms, the number of persons inhabiting each house 
being, therefore, much greater than is the case in this 
country. Each such " house " has a porter's lodge at the 
entrance, and it is outside this lodge that the notice 
previously mentioned is displayed, and it is in this lodge 
also that, at the pre-arranged time, the vaccinations and 
revaccinations are generally performed. For these opera- 
tions calf lymph is invariably employed, the process being 
carried out direct from calf to arm, a previously vaccinated 
calf being sent to the house from the Institut Vaccinal in a 
specially constructed van. Occasionally, from want of 
space in the porter's lodge or other reasons, the calf, 
after being removed from the van, is allowed to remain 
in the street, its halter being held by an attendant, while 
another assistant, taking his seat on a camp stool, pro- 
ceeds to collect lymph from the inoculated area of the 
animal's side and abdomen, with the aid of clamp forceps 


and lancet. Where such procedure is considered necessary, 
the persons requiring vaccination, whether infants or aduhs, 
also are brought out into the street, and the extraordinary 
spectacle may be witnessed of vaccinations being carried 
out by the medical staff surrounded by an interested crowd 
of sightseers. 

MM. Chambon and Menard attach much importance to 
this organisation and practice. They say that, under the 
immediate influence of existing smallpox, large numbers 
are willingly submitted, both to primary and secondary 
vaccination, who would otherwise escape ; and they are of 
opinion that certain classes who might object to be vacci- 
nated with lymph from an unknown source, find all their 
objections on this score removed when they actually see 
the calf which serves as vaccinifer. 

We were supplied both by Dr. Hervieux and by MM. 
Chambon and Menard with samples of the glycerinated 
lymph, which had been collected and prepared on the 
occasion of our visits. That which was obtained from the 
Academie de Medecine was collected on loth December 
1896. It was used by Dr. Cory at the Board's Animal 
Vaccine Station for the vaccination of twenty-seven children 
on 29th December. All the cases were, as usual, vaccinated 
by means of five insertions of lymph, and the insertion 
success obtained was 99.3 per cent. 

That obtained from MM. Chambon and Menard was 
collected on 8th December 1896 ; it was used by Dr. Cory 
on 22nd and 31st December, and on 7th January 1897, 
for the vaccination of ninety-six children, and every one of 
the five insertions succeeded in every child. 

[It should be stated, however, that these specimens of 
lymph obtained from Dr. Hervieux, and from MM. Chambon 
and Menard, respectively, were all found to contain large 


numbers of extraneous microbes when examined by the 
method of plate-cuUivation, even after the lapse of four 
weeks from the date of preparation. This fact is, no doubt, 
to be accounted for by reason of the percentage of glycerine 
employed not being sufficiently large. MM. Chambon and 
Menard informed us that they do not profess to attain 
bacteriological purity in the lymph distributed from their 
establishment. — S. M. C] 


At Brussels the propagation, storage, and distribution 
of calf lymph'is carried out at the Ecole Veterinaire, under 
the supervision of Professor Degive, the director of that 

The building set apart for the calf lymph station contains 
the director's room, a distributing room, an operating and 
preparation room, and two stables. As, however, we 
were informed by Professor Degive that the accommodation 
at present provided is regarded as very insufficient for the 
purpose, and that a new vaccinal institute is about to be 
erected, it would serve no useful purpose to enter into a 
detailed description of the present building. 

Stable. — The stable, which is a detached building, con- 
tain stalls for six calves, three on either side of a central 
footway, and the stalls are so arranged that a space is also 
left between them and the side walls of the building. The 
stalls are very narrow, and, at the end furthest from the 
central passage, have two iron uprights fastened to the sides 
of the stall. Iron rings, which are attached to the animal's 
halter by means of steel clips, slide up and down the 
uprights. This arrangement permits the calf to stand up 


or lie down, but prevents all possibility of its licking the 
inoculated portion of its body. 

The stable is warmed by an iron stove, the temperature 
at the time of our visit being 15° C. It is ventilated by 
windows opening inwards in the upper part of the two 
outside walls, and the removal of vitiated air is further 
facilitated by four outlet ventilators, each about 6 
inches square, and placed just above the floor level, the 
up-draught being aided by means of gas jets in the outlet 

Calves. — The calves are not of any special breed, and 
those that we saw did not seem to be so well suited for the 
purpose of lymph propagation as certain of those thus 
employed at the Institut Vaccinal in Paris. Another point 
of difference is that at Brussels male animals are used 
exclusively ; Dr. Degive believing that the finest vesicles 
are obtained on the scrotum. For the first four days after 
their reception the calves are kept in a separate " quaran- 
tine " stable. On the day prior to vaccination they are 
swung, by means of a belly-band and an arrangement of 
pulleys, into a wooden, zinc-lined, tank bath, capable of 
containing sufficient water to cover nearly the whole body. 
The temperature of the water is kept at about 30° C. In 
this bath the calves are scrubbed all over with soft soap. 
After removal from the bath, the skin is thoroughly dried 
with cloths, and the animal is then placed in the stable 
adjoining the operating room. 

Each calf is injected with 1 1 cubic centimetres of tuber- 
culin on the day prior to vaccination, but Dr. Degive 
considers this an unnecessary precaution for the reasons 
that {a) tuberculosis is very rare in calves, and that {b) no 
lymph is distributed until the animal from which it was 
obtained has been slaughtered, and necropsy has made it 


certain that the animal was not the subject of tuberculosis. 
In the event of tubercle being found, the lymph would be 

The age of the calves employed averages from ten to four- 
teen weeks, but animals four months old are sometimes used. 

The food of the calves consists principally of milk and 
eggs, each calf receiving, in twenty-four hours, 1 2 litres of 
milk and four eggs, together with a little hay, which is placed 
in each stall for the animal to eat if it is so disposed. 

Inoculation of Calves. — The calf is fixed, by means of 
ropes, to a tilting table of somewhat primitive construction, 
the right hind limb being elevated, as is usually done in 
England. A leather blinker is also fastened over the head. 
The right side and the abdomen are washed with soap and 
water with which lysol is mixed, and the surface is then 
shaved. The skin is afterwards washed with warm boracic 
acid solution, then with hot boiled water, and it is subse- 
quently dried with cloths which are sterilised by steam just 
previous to use. All instruments are also boiled in a 
solution of boracic acid. The lancets employed are similar 
in form to those devised by M. Chambon, while the com- 
pression forceps appeared to be of somewhat old design 
and needlessly heavy. 

The operator and his assistants all wear white blouses, 
which, just previous to use, have been sterilised in an 

Incisions, about 2 or 3 inches in length, are made 
at right angles to the long axis of the body, all over the 
shaved area of the skin, and also on the scrotum ; the 
average number for each calf being about 150. The 
incisions, which in each case are double, are made with a 
dry lancet, and are placed en eckelo?i, and about a couple 
of inches distant one from another. 


The lymph employed for vaccination is kept in stock 
for, at least, six weeks previous to its use ; it consists of vesi- 
cular pulp which, at the time of collection, is simply mixed 
with twice its weight of glycerine and is then kept in a 
glass tube, the mouth of which is closed with a cork fixed 
with paraffin until required. Just before it is needed for 
inoculation of a calf the pulp is ground up in a small agate 
mortar with a further small quantity of glycerine. The 
resulting emulsion is well rubbed into each separate incision 
on the skin by means of a thin ivory instrument resembling 
a small paper-knife ; the edge being passed up and down 
each incision several times. 

Collection of Lymph. — On the sixth day the calf is again 
fixed on the operating table, and the vaccinated area is 
once more washed with warm water or sterilised salt solu- 
tion and dried with sterilised cloths. The lymph required 
for stock purposes is then first collected. For this purpose 
compression forceps are applied to each vesicle separately, 
and the crust is first carefully removed with the edge of a 
lancet. These crusts are collected in a watch-glass, and 
are employed for the vaccination of children. The vesicular 
pulp is next removed by scraping with the lancet, and the 
material is collected in another watch-glass or Petri dish 
and weighed. Glycerine is added to it from a stock bottle 
to the extent of about twice the weight of the pulp, but the 
amount is only roughly estimated, no actual measurement 
or weighing being deemed necessary. The pulp and 
glycerine are stirred together, and are at once placed in a 
glass tube of such a size as to ensure its being almost 
entirely filled with the material available ; it is then fastened 
down by means of a glass stopper or cork, without further 

When sufficient pulp for stock purposes has been 


obtained, the remainder of the vesicles are scraped off with 
a Volkmann's spoon, and the material is mixed, as before, 
with glycerine, without trituration. 

The glycerine employed was stated to be of English 
manufacture, but was much thinner than that usually sold 
in this country, giving the impression, indeed, that it had 
been considerably diluted with water. It is sold as being 
" chemically pure." 

Storage and Distribution. — Just as is the case with what 
is termed the " stock " supply, this material is ground down 
in a mortar, with more glycerine, before being distributed 
for use. 

According to the amount required, the emulsion is either 
stored in tiny stoppered bottles, which are supposed to 
contain enough material for 25, 50, or 100 vaccinations; 
or, when a less quantity is desired, the material is placed 
in a slight excavation on the surface of a small glass plate 
about I inch square, and a plate of a similar size, but not 
hollowed out, is slid over it. The edges are sealed with 
paraffin, and the whole is wrapped round, first with tinfoil, 
and then with paper. 

The small bottles are fitted into blocks of wood, bored 
with holes for the purpose, in order that the parcel may go 
safely through the post. 

To each package is attached, by string, a doubled card, 
which can be addressed outside, and which, inside, has 
spaces for particulars as to number of vaccinations carried 
out, the number of insertions in each case, and the amount 
of success which results. 

The lymph which was supplied to us by Professor Degive 
was used for certain bacteriological investigations ; hence 
we have no record as to its success when used for the pur- 
poses of vaccination. 


by different vaccinators to Professor Degive, we found that 
these showed a high percentage of insertion success. 

While at Brussels, we also visited the Municipal Vacci- 
nation Station, of which Dr. Janssens, the Medical Officer 
of Health, is director. 

We were informed that the station, which consists of a 
waiting and an operating room, is open daily, but that, 
practically, no children are brought for vaccination during 
the winter months. This was unfortunately the case at the 
time of our visit, so that we had no opportunity of seeing 
the work in actual operation. 

The lymph employed is received in small glass-stoppered 
bottles from Professor Degive, of the Ecole Vdterinaire. 


^ The Animal Vaccine Establishment at Berlin, of which 
Dr. Schultz is the director, is situated in the Central Meat 
Market, on the outskirts of the city. 

The station consists of three parts connected with each 
other : ( I ) A large stable containing stalls for the calves ; 
(2) a work-room fitted with two tilting tables, somewhat 
similar to those in use in England, on which the calves are 
vaccinated, or the " lymph " collected ; and (3) the director's 
room, in which the lymph is triturated, glycerinated, and 
stored. This room contains cupboards and benches, and is 
fitted with all the necessary bacteriological apparatus, 
glassware, and instruments ; the latter being made entirely 
of metal, so as to admit of their being readily sterilised. 

Calves. — Cow-calves are almost invariably used, as less 
likely, when they lie down, to foul with their urine the 
vaccinated area of the abdomen than are males. The 
calves employed are usually from about six weeks to three 


months old. They are received forty-eight hours before they 
are required for vaccination, and are at once injected with 
half a gramme of tuberculin. If their temperature should 
rise above 41° C. during the next twenty-four hours they are 
not employed for the production of lymph. The calves are 
fed on a mixture of milk, eggs, and corn-flour, of which the 
milk is always sterilised prior to use. 

Vacci7tation of Calves. — When placed on the table, the 
abdomen is shaved from the vulva to the umbilicus, and a 
portion of the inside of each thigh is also shaved. The 
surface of skin thus exposed is carefully scrubbed with soap 
and water, washed with a solution of corrosive sublimate 
I- 1 000th, and then again washed with boiled water. The 
operator also washes his hands carefully, using carbolic 
soap, and before commencing to operate puts on a white 
cotton blouse over his coat. The calf s skin having been 
dried with a clean towel, long parallel incisions are then 
made over the whole length of the abdomen, and also over 
the shaved portion of the thighs. These incisions, which 
are made with a blunt knife, so as to draw as little blood as 
possible, are hardly a quarter of an inch apart, and are 
about 18 to 24 inches in length on the surface of the abdo- 
men. If any blood appears along the line of the incisions 
it is removed by means of sterilised blotting-paper. A few 
grammes of stored glycerinated lymph, prepared some 
weeks or months previously, are next poured on the abdomen 
and spread over the incised lines with the back of a scalpel. 
Collection of Lymph. — On the fourth or fifth day (96 
or 120 hours after vaccination) the calf is again placed 
on the table. After a thorough cleansing of the skin in 
the same manner as before, absolute alcohol is poured 
over the vaccinated area. When the alcohol has evaporated 
the surface is treated with ether, which is supposed to exert 


a bactericidal, in addition to its anaesthetic action. The use 
of alcohol and ether has, however, been discontinued since 
the date of our visit, soap and water merely being now 
employed. Then the skin is put on the stretch and 
scraped, in the direction of the incisions, with a rather blunt 
" Kartoffel Loffel." This spoon is taken over each portion 
of the vaccinated surface once only, so as to avoid, as far 
as possible, admixture of blood ; and by this means all the 
epithelium which has undergone vesicular changes, caused 
by the action of the specific virus, is removed in long strips 
of about one-eighth inch wide. Compression forceps are 
not needed, and the whole operation is completed in a few 

Preparation of Lymph. — The whole mass of epithelial 
tissue removed by the spoon is collected and emptied into 
a glass Petri dish, and afterwards it is weighed in a delicate 
balance. Seven times the weight of cold boiled water 
and a similar quantity of glycerine are then weighed out 
separately. A small portion of the water is added to the 
dish containing the tissue scrapings, and after being stirred 
together the mixture is passed between the small porcelain 
or glass rollers of a mixing mill, invented by Dr. 

Description of Dr. Doering's Lymph-Grinding 

The apparatus consists of an iron frame, resting on a 
metal base, and carrying four unsealed glass rollers. 
These are easily removed and sterilised, but care is 
taken that each roller is replaced in its original position. 
The spindles lie in slots, and by means of springs the 
rollers are pressed against each other. The rollers are 


made to revolve by cog-wheels attached to the spindles 
on the exterior of the frame. The whole system is set in 
motion by turning the wheel No. 3 with the handle 
(which is screwed to its spindle) in a direction from 
the operator. 

The mixture, lymph -pulp and glycerine, on being 
poured between the upper two rollers soon becomes dis- 
tributed over the lower system as well. After it has 
reached roller No. i, it is scraped off by a glass plate 
placed below, the edge being pressed against the rollers 
by a spring. From the plate the lymph emulsion flows 
into a square porcelain dish provided with a spout. To 
prevent the lymph spreading to the ends of the rollers, 
glass guides are attached to rollers Nos. 2 and 3, which 
conduct the emulsion towards the centre. 

The method of working is as follows : — 

After loosening the grips, right and left, fold back both 
arms and remove rollers Nos. 3 and 4. Unscrew both 
glass guides and lift out the bridge together with rollers 
Nos. 2 and i. Rollers, glass guides, and glass plate are 
then sterilised, which process may be accomplished without 
withdrawing the spindles or removing the glass guides 
from their clamps. 

The apparatus can be put together again in a few 
minutes. To do this, first replace the glass plate and then 
rollers Nos. i, 2, and 4. Next press roller No. 3 gently 
back into position. Then fold over both arms, and finally 
fix the bridge and guides by means of the side-screws. 

The mixture of lymph-pulp and glycerine is now poured 
little by little between rollers Nos. 3 and 4, the larger 
shreds of epithelium being taken up and placed in position 
by means of forceps. A second person turns the handle, 
and, in a short time, the mass, now more or less finely 


Lymph-mixing machine of Dr. Doring, Berlin. 


triturated, appears on roller No. i, from which it flows 
into the porcelain dish placed beneath. After the whole 
quantity of material has passed through, the resulting 
emulsion is stirred with a sterilised glass rod, more 
glycerine is gradually added, and the mixture is then 
passed a second and, if considered necessary^^ a third time 
through the machine. The lymph after such further 
passage through the mill forms a homogeneous emulsion, 
the individual particles in which are of exceptional 
fineness. '• 

The loss of material during preparation is exceedingly 
little, and the working up of the produce of a calf, amount- 
ing to about 20 grammes, can be completed in twenty 

Formerly, it was the custom at this station to add both 
glycerine and water to the epithelium partly before and 
partly during its passage through the mill. 

For some months past, however, it has been the 
practice not to add the glycerine until after the material 
has been twice passed through the mill. This grinding 
process is eflfected with more difficulty in the absence of 
glycerine ; but the reason for the alteration is that much 
of the lymph is now centrifugalised,i a method of pro- 
cedure which would be unduly prolonged if the specific 
gravity of the emulsion operated on had previously been 
increased by the addition of glycerine. The centrifuge at 
present in use is a small two-armed instrument worked by 
hand ; its use involving the employment of an extra assist- 
ant for at least a couple of hours. At the end of this time 

1 This process, which was only tentative, has now (1898) been in 
great measure abandoned. 


the minute shreds of epitheHum contained in the mixture 
have settled into a compact mass at the bottom of the tube, 
and the supernatant fluid is only very slightly opalescent. 
This is decanted off, and an amount of glycerine equal in 
weight to that of the water previously employed is in- 
timately mixed with it, after which the resulting " lymph " 
is stored in a stock bottle fitted with an indiarubber 
stopper and cap ; or it is put up in small glass tubes of 
I cc. capacity, each of which contains, according to Dr. 
Schultz, sufficient material for loo vaccinations. The 
amount of vesicle pulp collected from a single calf varies 
from lo to 15 grammes. This, when intimately mixed 
with the usual amount of dilute glycerine, is calculated to 
provide sufficient material for the vaccination of, at least, 
I 5,000 persons. 

This process of centrifugalisation is as yet only tentative 
and experimental. The appearance of the " lymph " is 
thought to be improved by its adoption, and, when tested 
by the method of plate cultivation, it is found to be freer 
from " extraneous " microbes than is an equivalent amount 
of the emulsion when tested before treatment in the centri- 
fuge. Objection, however, to the employment of the 
method migh^ be based on this freedom if, as there is 
every reason for beHeving, the microbe specific to vaccine 
is present in a far greater amount within the cells of the 
vaccinated dermis than in the intercellular lymph spaces. 
Even if free in the fluid portion of the mixture, and of 
exceptionally minute size, the continued action of the 
centrifuge must tend in time to remove them, just as is 
found to be the case with other microbes which may be 

The amount of glycerine and water employed in the 
preparation of vaccine material has been considerably 


decreased during the past twelve months, the relative pro- 
portions being at present : — 

Epithelial pulp . . . . i part 

Glycerine ..... 4 parts 

Boiled water .... 4 parts 

All " lymph " is nov^^ tested bacteriologically by means 
of plate cultivations, before being distributed. This is 
done in consequence of the recommendation of a scientific 
committee of which Professor Koch was a member and 
which has recently been sitting at Berlin to inquire into the 
whole subject of the collection, purification, and preserva- 
tion of vaccine lymph. 

Season of Calf Inoculations. — Inoculations are only 
carried out in the months of May, June, and July. The 
calves being themselves vaccinated with stored glycerinated 
lymph, it is not necessary to keep going a continuous 
series ; and in these three months sufficient lymph is 
manufactured for use during the whole year, throughout 
one of the largest of the eight districts into which the 
kingdom of Prussia is divided for vaccination purposes. 

Disposal of Calves. — After collection of lymph, the 
calves are sold to the Jewish Rabbi to be slaughtered for 
food. We were informed that a larger price is given for 
them than is ordinarily the case with calves brought to the 
Central Meat Market, owing to the fact that they are in 
such fine condition as the result of good feeding while at 
the station. 

Glycerinated calf lymph, collected and prepared as above 
stated on the occasion of our visit on loth January 1897, 
was used by Dr. Cory for the vaccination {a) of thirty 
children on 19th January, with an insertion success of 97.1 
per cent; {b) of six children on 21st January, with an 


insertion success of 86.6 per cent; {c) of forty children on 
25th February, with an insertion success of 92.3 per cent ; 
and of thirty-three children on 4th March, with an insertion 
success of 67.5 per cent. This sample of glycerinated 
lymph was again used after having been kept until six 
months had elapsed since the date of its preparation. It 
was then found that its activity as vaccine lymph had 
practically disappeared. It is right to state in this connec- 
tion that at the Board's Animal Vaccine Station there are, 
as yet, no means of storing lymph elsewhere than in the 
somewhat high temperature at which the operating rooms 
are maintained. 


The Animal Vaccine Institute, of which Dr. Chalybaus 
is director, is situated in the northern suburbs of Dresden. 
It consists of a small two-storied building, containing, on 
the ground floor, an operating room and three other rooms, 
while the whole of the first floor is utilised as a dwelling for 
the caretaker. Adjoining this building is a small stable 
containing stalls for the calves. 

Dr. Chalybaus informed us, however, that the Animal 
Vaccine Institute was hardly arranged in accordance with 
modern requirements, having been established more than 
twelve years ago, when the present methods of preparing 
lymph were not in vogue. 

The stable in which calves are placed on arrival contains 
two stalls and a tank bath in which the animals are 
thoroughly washed before use. The calves are lifted into 
and out of this tank by means of belly-bands attached to a 
system of pulleys fixed to the ceiling. After having been 
dried with cloths they are vaccinated and then placed in 
another stable on the opposite side of the house. 


The calves are bedded in their stalls on fine wood 
shavings, which are said to have the advantages of being 
clean, dry, and comfortable. 

The operating room is about 20 feet square, and 
contains two tables. One of these is for calves ; the other, 
of larger size, and fitted with mechanical arrangements for 
tilting and raising, is for young bullocks, which are 
occasionally employed for purposes of vaccination, when 
either an extra amount of lymph is required or when it has 
been impossible to obtain the required number of calves. 

By preference. Dr. Chalybaus employs cow-calves of 
from six to eight weeks old. 

The calf table is an oblong, shallow trough of wood, 
provided with straps and with two iron uprights at one end, 
to which the hind limbs of the calf are fixed in a V-shape. 
This method of fixation, however, enables the animal to 
struggle to such an extent as to raise its hind-quarters com- 
pletely off the table. 

After having been shaved, and before vaccination, the 
animal's skin is washed with soap and hot water containing 
lysol. The soap suds having been washed off with more 
water, and the skin dried with a cloth, benzine is poured 
over the surface to render it more aseptic, and is rubbed 
in with sterile sponges of gauze, which are kept for use in 
a sterilised glass-stoppered bottle. Lengthy incisions are 
next made with a blunt scalpel, in the long axis of the 
body, over the inside of the thighs and over the whole 
surface of the abdomen from the vulva to the umbilicus ; 
also over the lower ribs. Glycerinated lymph which has 
been stored in sealed tubes for from three to as long a period 
as eighteen months is next rubbed in over the area of the 
incisions with the flat surface of a small trowel-shaped 


We were informed that much difficulty is experienced in 
this establishment in obtaining calves suitable for purposes 
of vaccination, as throughout Saxony it is the custom to 
slaughter these animals for food at a very early age, some- 
times within a few days of birth. This being so, calves 
have to be imported from a distance, most of those employed 
by Dr. Chalybaus coming from Berlin or Hamburg. They 
are obtained by a local cattle-dealer, who charges twenty 
marks for their use, and who removes them for slaughter 
immediately after collection of the lymph. 

Calf-to-arm vaccination is never employed in Saxony, as 
it is thought to be undesirable to use lymph from an animal 
until a necropsy has shown it to have been entirely free from 

Collection of Lymph. — The lymph, or rather the vesicle 
pulp, is collected after an interval of four complete periods 
of twenty-four hours. The skin is first washed with white 
soft soap and hot water, the operation being carried out 
with the aid of a large house-painter's brush. Such crusts 
as have formed are removed as far as possible with the 
edge of an ordinary metal teaspoon, after which glycerine 
is poured over the skin and rubbed in with gauze 

The pulp is collected by scraping with a Volkmann's 
spoon, but as Dr. Chalybaus goes over the same surface 
again and again, a not inconsiderable amount of blood 
becomes mixed with the epithelial scrapings. The raw 
surface of the abdomen is afterwards dusted over with fine 

The pulp thus collected is weighed and is then run 
through a mixing machine invented by Dr. Chalybaus, of 
which a special description, written by him, together with 
an illustration, is appended to this report. The necessary 


motive power is supplied from a small water motor fixed 
beneath the floor of the room. 

After being ground up in the machine, four times the 
amount of a mixture of glycerine and sterilised water (water 
3 parts, glycerine i part) is added to the vesicle pulp, and 
the whole is then run through once again to ensure thorough 
admixture. The resulting emulsion is received into a 
porcelain mortar placed beneath the machine. The mortar 
is removed when all the material has passed through, and 
its contents are then taken up by suction into tubes of 
somewhat large calibre which, when filled, are closed at 
either end by means of sealing wax. 

Dr. Chalybaus considers that i gramme of glycerinated 
emulsion is sufficient for 80 vaccinations. The vesicle 
pulp obtained from a single calf affords from 50 to 75 
grammes of the glycerinated emulsion ; or, in other words, 
enough for the vaccination of from 4000 to 6000 persons. 

Glycerinated calf lymph, collected and prepared by Dr. 
Chalybaus on the occasion of our visit on 12th January 
1897, was used by Dr. Cory for the vaccination of 15 
children on 2 1 st January. The children were, as usual, 
vaccinated by means of five insertions, and every insertion 
gave a successful result. 

[A further sample of lymph collected on 12th January 
1897 was employed by Dr. Cory nearly six months later, 
for a further series of vaccinations. As a result he again 
attained complete insertion success. The lymph examined 
bacteriologically at this date by the method of plate 
cultivation was found to be completely free from extraneous 
micro-organisms. — S. M. C] 



Report of the Royal Institute for Vaccination 
IN Dresden. 

About the Technique of Preparing Animal Lymph ^ 
by Dr. Chalybdus, Dresden. 

As a rule, the exclusive use of animal lymph for vaccina- 
tion has been everywhere adopted, whilst the use of human 
lymph, taken from children, has been almost entirely 
abandoned, and public institutes for vaccination prepare 
only calf lymph. The preparation of animal lymph needs 
special contrivances, because the lymph scraped out of the 
" smallpox of a calf," in order to make it available, must be 
triturated and ground to a fine pulp, and with the addition 
of glycerine, turned into a thin and homogeneous emulsion. 
When triturated in a dish or bowl, this operation takes, for 
the quantity of lymph obtainable from one calf, three hours' 
time, and cannot therefore be done by the physician himself ; 
nevertheless, great care must be taken, as it is unwise to 
leave that part of the preparat?bn of lymph to a common 
workman, who cannot be properly and continuously watched. 

As far back as 1889 I constructed a machine for 
triturating lymph, which stood the test of use in the Institute 
of this country and of those of many others. In 1893 the 
machine was improved. It is now fitted upon the marble 
table of a sewing engine, and moved either by foot or, still 
better, by means of a small hydraulic, steam, or electric 

A cylinder formed out of two equal parts, having in its 
inside threads of a screw, is attached to the shank of a 
column ; a close-fitting spindle, likewise provided with 
threads of a screw, turns in the cylinder. Through a 


Lymph-mixing machine of Dr. Chalybaus, Dresden, as modified for use 
in the laboratories of the Local Government Board. 


funnel the raw lymph is put into the upper end of the 
cylinder ; in turning the spindle the lymph is rubbed and 
ground to the utmost, and leaves the cylinder on the lower 
opening, dropping into a glass dish. 

By means of this contrivance the preparation is com- 
pleted in about fifteen minutes, although the work is done 
in a far better way than as usually in a dish.' The lymph, 
consisting of coherent and liquid parts, is thoroughly rubbed, 
not only squeezed, and turned into homogeneous emulsion. 
It does not lose its natural colour, nor does it get warm. 

The apparatus is made of steel bronze, and its parts, 
comprising cylinder, spindle, and funnel, can easily be 
separated and disinfected in boiling water or otherwise. 

The trituration occurring whilst the apparatus is closed — 
the opening of the funnel is closed by a cover of glass — no 
dust can enter, and should a calf hair accidentally have 
dropped into the lymph it can be removed whilst the lymph 
slowly drops out of the cylinder. No loss of lymph can 
occur, because the machine retains none of the material 
with which it is fed. 

The machine can be obtained through the Royal Institute 
for Vaccination, in Dresden, price 200 marks. Every 
machine is tested by the president of the above-mentioned 
institute, and a certificate is granted to each one sent out, 
as a proof of its fitness. 

Directions for using the Machine for triturating Lymph^ 
invented by Dr. Chalybdus 

I. To take the jnachine to pieces^ slack the screw in claw 
/on which the driving-wheel g and spindle c are attached. 
Open the claws and remove the spindle and driving-wheel 
g^ taking care not to let the former drop. Take off funnel 
^, slack screw of lower clamp d^ and unscrew the middle 


clamp e^ holding the cylinder in the left hand. Take the 
cylinder out of the support h. In order to separate the 
two parts of cylinder, use a small piece of wood by inserting 
it in the hollow, but care must be taken not to drop the 
parts when separated. Remove the driving-wheel from the 
spindle, and clean it, as well as the spindle and funnel, with 
a brush, and steriHse in boiling water. 

2. To put the 7nachine together^ first join both parts of 
the cylinder, put on the lower clamp and turn its screw a 
little, pass the cylinder vertically through the support, and 
place the funnel on the top of the cylinder. Then screw 
the driving-wheel on the upper part of the cylinder, open 
the claws and insert the spindle so far that the claws grasp 
the spindle close under the driving-wheel, shut the claws, 
and screw tight. Bring the lower ends of spindle and 
cylinder exactly in a line and fasten the lower clamping 
screw. Finally, adjust the belt on the driving-wheel. 

3. To put the machine in vwtioti^ move the fly-wheel k 
outwards, and to avoid friction use vaseline for lubricating 
the moving parts. It is advantageous first to run some 
glycerine through the cylinder, after which the lymph is 
placed in the funnel and the cover adjusted. If preferred, 
however, glycerine can be added to the lymph, and the 
mixture passed through the machine. The finished lymph 
leaves the lower end of the cylinder and drops into a small 
glass vessel /. 


The buildings provided for this Institution are the most 
recent of their kind in Germany, and, as we were informed 
previous to our visit, all the fittings are of the most modern 

We visited the Institution with Dr. Vanselow, where we 


were also met by the assistant director, the veterinary surgeon 
attached to the staff, and certain other gentlemen. 

Dr. Vanselow presented us with a reprint of a paper 
written by him, showing a description of the buildings, 
illustrated with blocks giving the elevation and ground 
plans.i We append a translation of this, so that it is 
unnecessary here to enter into any details as to structure. 

Stable. — The calf stable, which adjoins the collecting 
room, contains ten stalls, one of which is reserved for any 
calf used for experimental purposes, while another is merely 
a pen forming the platform of a weighing machine. The 
sides and ends of the stalls are formed solely of a series of 
iron bars, painted grey, and they are of such narrow width 
as to make it impossible for a calf to turn round, it being 
thus prevented from licking the inoculated area of its 
abdomen. Each stall is provided with a gate at either end, 
opening outwards, and bears a numbered label of iron. The 
flooring of the whole stable is formed of cement concrete, 
over which in each stall is placed a wooden rack or platform 
which is raised about 3 inches from the floor. These 
racks are formed of wooden splines about 2 inches square, 
placed close together, the upper edges of each spline being 
slightly rounded. On these racks the calves stand or recline, 
no bedding of hay or straw being employed. 

Calves. — The calves range usually from about six to 
eight weeks old. They are purchased in the meat market 
which immediately adjoins the establishment. The calves 
required for the current week are bought on Monday, and 
they are sold on the following Saturday, after their slaughter 
and the collection of lymph on the previous day ; so that 
the stable is always empty from Saturday to the following 
Monday. The calves are kept under observation in the 

1 Plans not reproduced here. — S. M. C. 


stable for twenty-four hours after their reception, and are 
vaccinated on Tuesday. Only three complete days are 
allowed for the progress of the local results of vaccination. 
On Friday the animal is slaughtered in a small slaughter- 
house opening off from the stable, and immediately it is dead 
the carcase is brought into the collecting room on a trolley, 
the abdomen washed, and the epithelial pulp of the vaccina- 
tion area is removed by means of a sharp spoon. 

Vaccination of Calves. — We did not see the process of 
vaccinating the calves, but we learnt that it was carried 
out in an exactly similar fashion to that employed at the 
Berlin and Dresden stations, namely, by long parallel 
incisions over which glycerinated lymph is rubbed by 
means of a spatula or other flat-bladed instrument. The 
lymph employed for the vaccination of calves is always 
kept for a period of at least six weeks after glycerination, 
in order to ensure that it shall be as free as possible from 
extraneous organisms before it is used to vaccinate the 

After collection of the lymph pulp, and while it is being 
prepared for use, the carcase of the calf is taken back to 
the slaughtering room, where it is skinned and opened. 
The internal organs are removed and brought in on trays 
to be examined by the veterinary surgeon. In the event 
of his forming the opinion that any of the organs presented 
any condition indicative of disease, the lymph derived from 
the animal in question would be at once destroyed. 

In view of this precautionary measure it is not deemed 
necessary to test the calves by the injection of tuberculin 
prior to their vaccination. 

Collection and Preparation of Lymph. — The greatest 
amount of vaccine is collected during the months of March, 
April, and May, when from six to eight calves are employed 


every week. For the remainder of the year the weekly 
vaccination of one or two calves is found to be sufficient to 
supply all the lymph required for human vaccinations and 
revaccinations in the Cologne district. 

In the preparation of the lymph material the epithelial 
pulp from the vaccinated area is removed by scraping with 
a Volkmann's spoon, and is received in a small glass dish. 
In this it is weighed, after which it is turned out into a 
mortar and thoroughly triturated ; at first without any 
addition of water or glycerine ; later, small quantities of 
water are gradually added to the extent Qi Jive times the 
weight of pulp. The mixture having been ground up still 
further, double the quantity of glycerine is finally incor- 
porated. Thus, at the time of our visit, 10 grammes of 
pulp having been collected from one calf, the composition 
of the finished emulsion was as follows : — 

Pulp . . « .10 grammes 

Water .... 33.5 
Glycerine .... 66.5 ,, 

Dr. Vanselow informed us that this quantity would 
suffice for the vaccination of 5000 children. 

The emulsion is afterwards forced into small bottles and 
tubes by means of a machine actuated by water power. 
This machine, which is manufactured by a Vienna firm, 
appears to be decidedly useful and convenient, and is 
capable of being worked in connection with any form of 
pressure apparatus. 

The small bottles are of different sizes, and contain 
lymph sufficient for 50, for 100, and for 150 vaccinations 
respectively. The bottles and their corks are all of them 
sterilised prior to use. 

We were struck with the numerous precautions which 


are taken in this institution to ensure thorough asepsis 
throughout the various stages of lymph production. Thus, 
in the collecting room, the flooring is of cement concrete^ 
the walls are lined internally for about half their height 
with opaque glass tiles, the upper half being of Parian 
cement. The shelves are of glass supported on iron 
brackets, and the surface of all tables consists of thick 
slabs of glass over green cloth, the glass being removable 
if necessary. The institution is furnished throughout with 
electric light, and sterilised hot water is supplied as needed 
from a small apparatus affixed to the wall of the collecting 
room. Indiarubber pipes used in connection with the hot- 
water apparatus and the pressure apparatus employed for 
filling tubes are kept in a strong solution of carbolic acid 
when they are not in use. The director and all his 
assistants wear linen covers over their clothes, but only the 
sleeves, which are detachable, appear to be sterilised prior 
to each occasion on which they are used. 

On the first floor of the building is a bacteriological 
laboratory, which is reached from the collecting room by an 
iron spiral staircase. It is fitted up with an autoclave, 
incubators, etc. But owing to the director being engaged 
in private practice, he has no time to work in this laboratory, 
which is, therefore, only used when it is desired to sterilise 
cloths, instruments, or glass-ware. 

With the lymph and pulp material collected and 
glycerinated in our presence on i6th January 1897, Dr. 
Cory vaccinated thirty-four children on 19th January, with 
an insertion success of 98.8 per cent, and twenty-one 
children on nth February, with an insertion success of 93.3 
per cent. 


( Translation) 

The Royal Lymph Station for the Rhine Province 
AT THE New City Cattle and Slaughtering 
Establishment, Cologne, by Sanitary Coun- 
cillor, Dr. Vanselow, Director. 

The public institution for the preparation of animal 
lymph to meet the needs of the Rhine Province and the 
Hohenzollern districts was erected in Cologne in 1889. 
The rooms at the old slaughtering establishment which 
were used until 1895 were extremely defective, being 
narrow, damp, and dark, and rendered any practically 
uniform arrangement impossible. In building the new 
cattle and slaughtering establishment, the provision of a 
suitable annexe for the production of lymph was borne in 
mind from the very first ; and so the present institution 
originated, which meets all the demands of hygiene, and 
may justly be regarded as the prototype of such institu- 

The annexe is situate at one end of a large cattle shed, 
both having a common partition. The main front faces the 
north, so that all the rooms, as there are windows only in 
the front, receive their light equally distributed from that 
direction. The building is very solidly built, but under- 
cellared only, and to the smallest extent, on the eastern 
side. As, however, the whole of the ground was filled in, 
and only absolutely dry and permeable material employed, 
there is no fear of any dampness of the rooms. 

On the ground floor of the building are the corridor, 
collecting room, calf stable, slaughter room, doctor's room, 
office, and a closet. 

On the first floor, which is reached from the collecting 


room by a convenient winding staircase, is the laboratory, 
adjoining which, on both sides, is a large garret. The 
entrance to the building is from the street, and so arranged 
that the cattle establishment has not to be traversed in 
order to enter ; the wall belonging to the entrance being 
continuous with the wall which encloses the cattle estab- 
lishment. The calves are driven into the station through 
the doorway of the slaughtering room, which lies at the 
opposite end of the building. 

While the floors of the slaughtering room, calf stable, 
collecting room, and laboratory are of concrete, the doctor's 
room and the office have inlaid wooden floors (parquetry) ; 
the corridor and closet are laid with "Mettlacher" tiles; 
and, finally, the rooms in the roof (garrets at the sides of 
the laboratory) are laid with floor boards. In the 
slaughtering room, calf stable, and collecting room the 
floor is slightly sloped in one direction, and at the lowest 
point there is a drainage outlet which is shut off by a small 
intercepting trap. The height of the rooms on the ground 
floor is 3 J metres, except the collecting room, however, 
which is 4j metres in height, and that of the laboratory is 
3 J metres. All the rooms on the ground floor are vaulted 
with plain solid arches. The laboratory and garrets have 
rafters and wooden ceihngs. The collecting room, calf 
stable, and closet are lined, to the height of \\ metres from 
the floor, on all four sides, with white opaque glass tiles ; 
the slaughtering room, on three sides (the door side is 
excluded), to the same height with white glazed tiles. The 
remaining part of the walls in these rooms is painted with 
white porcelain enamel. The walls in the doctor's room 
and the office are papered. In the laboratory and corridor 
the walls are painted with oil paint. The collecting room 
is brightly lighted by a window 4 metres wide by 3 metres 


high. The light entering becomes strongly reflected by 
the brilliant white walls. The laboratory has three 
windows, the calf stable two, and the doctor's room and 
•office one large window each. The closet and slaughtering 
room have each one small window. All the rooms are, 
therefore, amply lighted. 

The calf stable contains eleven stalls for the reception of 
the calves, i.e. six on one side and five on the other side of 
the centre passage. The stalls are so constructed that the 
wall nowhere forms the boundary of a stall, and a clear 
passage is thus given all round the stalls ; they are each 
70 centimetres wide and 150 centimetres long. The 
enclosures are formed of iron lattice - work ; at the two 
narrow sides of each there is a door permitting the calves 
to be taken in or out by either side according to con- 
venience. The iron lattice -work is painted a light grey 
colour, so that every speck of dirt can at once be seen and 
removed. On the floor of the stalls lie wooden gratings. 
Upon the space which would correspond to the twelfth 
stall a weigh -bridge is sunk, enclosed with the above- 
described iron work ; this machine allows of the calf being 
weighed while being taken through. The hollow in which 
the weigh -bridge stands has also a smell -preventing 
arrangement. The ventilation of the stable is obtained by 
a large tube which passes through the garret above. The 
collecting room has a flap ventilator in the window, as 
shown on the plan. Between the calf stable and collecting 
room there is an arrangement of double doors, one of which 
is thickly padded. The doors, padding, and through-air 
draught isolate the collecting room both from smell and 
noise. The closet is a so-called " Unitas " closet. Water 
is supplied by the city main, and in every room a sink is 
provided. The lighting is by electricity ; in the laboratory 


however, the " Au^rches Gliihlicht " (a form of incandescent 
light) has been found preferable. 

The heating is effected by means of American stoves ; 
for the doctor's room, however, a gas stove has been pro- 
vided by reason of its greater suitabiHty. Large gas stoves 
serve for the heating of the water and milk. For the 
storage of considerable quantities of lymph a sufficiently 
large room is reserved in the refrigerating house of the city 
cattle establishment. 

The furniture provided for the institution is worthy of 
the handsome rooms, and consists throughout of oak. The 
laboratory is completely fitted for bacteriological investiga- 
tions, containing all sterilisation apparatus — thermostats, an 
excellent microtome, microscopes, centrifuge, etc. 

The extent of the lymph production may be understood 
from the following figures : — In the year 1 894 about 356,000 
portions of lymph were issued, and in this year (1895) the 
number of portions will nearly reach 400,000. 


The Institut Vaccinal Suisse, which was visited by one 
of us (Dr. Copeman) only, is situated at Lancy, on the out- 
skirts of Geneva. It was founded in 1882 by M. Charles 
Haccius, the present director of the establishment. 
Originally a private venture, it is now recognised by the 
various cantonal Governments, M. Haccius, in considera- 
tion of an annual subvention, supplying to public vaccinators 
throughout Switzerland, free of cost, all the lymph required 
by them in the performance of their duties. 

The building in which the Vaccine Institute is housed 
adjoins a model dairy, also established and carried on by 
M. Haccius. The institute building contains two stables 


for calves, an operating and collecting room, a laboratory, 
a room in which the packing and distribution of the lymph 
are carried out, and the director's room. 

Stables. — Each of these contains four stalls. The side 
walls of the reception stable are of concrete ; those of the 
stable adjoining the operating room are of wood. The 
floors of both stables are of concrete. The stables are 
kept at a temperature of 18° C. to 20° C. Calves are 
received into the first-mentioned stable, and are there kept 
under observation for four or five days, at the end of which 
period they are passed into the stable next the operating 
room. The bedding of the stalls consists of fine wood 
shavings, this material being, in the opinion of M. Haccius, 
decidedly preferable to straw as regards both the cleanliness 
and comfort of the calves. 

Calves. — These range usually from about three to four 
months old. They are fed on milk, obtained direct from cows 
in the adjoining dairy, in addition to which they are allowed 
oatmeal and a certain number of eggs, but no hay. The calves 
are purchased from peasants in the surrounding districts. 
After vaccination and collection of the lymph, they are sold 
to a butcher in Geneva, at a loss of about ^i on each calf. 
They are slaughtered in the public abattoir, and the veterinary 
inspector attached to that establishment furnishes a certificate 
relating to the healthiness of the calf and the condition of 
the viscera as ascertained on examination of the carcase. 

Vaccination of Calves. — For the purpose of vaccination 
the calf is strapped down to a tilting table, similar in its 
main features to those employed in England. The head of 
the animal is covered with a leathern mask. The whole of 
the abdomen, the inside of the thighs, and a considerable 
area of the right side of the body of the animal are shaved, 
white soft soap and hot water being used in the process. 


The skin is next washed with solution of lysol (2. 5 parts per 
1000), and finally with hot boiled water. It is dried with 
sterilised gauze sponges. The actual vaccination is carried 
out in a manner similar to that universally employed in the 
German Government establishments already described, with 
the exception that the parallel lines of incision are discon- 
tinuous at intervals of about four inches. Occasionally a 
certain number of incisions are madeat a greater distance from 
each other, and only about a couple of inches in length, in 
order that the condition of the resulting vesicle may be more 
readily observed. Any blood which exudes from the incisions 
is removed with sterilised gauze sponges, and then the skin 
is put on the stretch, while glycerinated lymph is rubbed into 
each incision by means of a small and thin ivory spatula. 
The lymph employed consists of one part of vesicle pulp 
incorporated with two parts of undiluted glycerine, and the 
resulting mixture is stored for about a month prior to use. 

Collection and Preparatiofi of Lytnph. — After the lapse 
of four days and a half from the time of vaccination, the 
calf is again placed on the table, and the vaccinated area 
washed with warm boiled water without the addition of any 
antiseptic. After drying with sterilised gauze sponges, the 
vesicle pulp is removed by scraping with a sharp spoon. 
The resulting pulp is collected in a glass pot provided with 
a cover, and when all has been removed the total amount 
is weighed. Sufficient glycerine (undiluted) is then added 
to cover the mass of pulp, and the vessel and its contents 
are set aside for a few days. Subsequently, glycerine and 
water are added in proportions requisite for attaining the 
following standard : — 

Vesicle pulp . . . .1 part 

Glycerine . . . . .2 parts 

Water . . . . .1 part 


and the mixture is then thoroughly triturated in a mixing 
machine of the kind invented by Dr. Chalybaus of Dresden. 
The resulting emulsion is employed for human vaccinations, 
the " seed material " used for the vaccination of calves, 
having, as already stated, no water added to it. 

Occasionally clamp forceps are employed in the collec- 
tion of lymph from the smaller vesicles, when it is required 
to store it along with glycerine in fine capillary tubes ; the 
resulting material, containing comparatively little epithelial 
tissue, being therefore more readily drawn up into the tubes. 
When collection is carried out in this manner the " crust " 
is first removed from the vesicle, which is then gently 
scraped with a lancet. The material thus obtained is 
mixed with glycerine in the usual fashion. 

The emulsion, which is never sent out for use until at 
least four weeks after collection of the vesicle pulp, is 
stored prior to distribution in large glass-stoppered tubes. 
It is sent out in flacons, plaques, and capillary tubes, 
according to the amount required in any given case. The 
flacons, small glass tubes made of amber- coloured glass 
and provided with corks, are of sufficient size to contain 
enough emulsion for 25, 50, and 100 vaccinations respec- 
tively. The plaques consist of two small squares of glass, 
one of which has a shallow excavation on one surface. 
This is filled with emulsion, then covered with the plain 
square, and the edges sealed with paraffin. Quantities of 
emulsion sufficient for five or ten vaccinations are sent out 
in this way ; while fine capillary tubes, which are sealed 
with paraffin, are used for sending out lymph for the vacci- 
nation of one person only. All flacons, plaques, and tubes 
are sterilised before being filled. In order to send them 
safely through the post they are enclosed in neat metal 
cases diffisring in size and shape. These, together with 


certain printed matter, including a card to be filled up in 
accordance with the results obtained from use of the con- 
tained lymph, are enclosed in a stout glazed orange-coloured 
envelope secured with a metal clip. 

M. Haccius stated that he had, especially of late, 
experienced some difficulty in getting public vaccinators to 
fill up and return the cards sent with each consignment of 
lymph, so that it was not possible to obtain full statistics as 
to the success attending the use of the lymph sent out from 
the Institute. On looking over with him, however, a 
number of cards which had come to hand within the last 
few months, it appeared that in all cases the success 
attained was very great ; in a not inconsiderable proportion 
the insertion success had reached loo per cent. 

M. Haccius further stated that the structural arrange- 
ments of his Institute were the same as when it was first 
started in 1882, so that they are not in some respects such 
as would be considered most desirable at the present time. 
Nevertheless, it was impossible not to be impressed with the 
strict precautions taken to ensure the utmost cleanliness in 
the case both of the premises and of all persons employed 
in the various details of the work. 



Extracts from the Transactions of the Seventh 
International Congress of Hygiene and 
Demography: London, 1891. Vol. II. Section 
II. pp. 319-26. 

The Bacteriology of Vaccine Lymph : by S. Monckton 
Copenian^ M.A,, M.D. Cantab.^ Research Scholar 
of the British Medical Association^ late Assistant 
Lecturer on Physiology^ St. Thoinas's LLospital. 

Numerous have been the attempts, ever since bacteriology 
first took rank as an exact science, to solve the problem as 
to the nature of the active principle of vaccine lymph, and 
in consequence of this, and of the great difficulty of the 
subject, a mass of literature has gradually accumulated 
sufficient to appal any one newly entering upon the task. 

The fact, however, that in the absence of any sufficient 
knowledge on this point, the whole practice of vaccination 
as at present carried out is more or less empirical, may 
perhaps furnish excuse for still further attempts in this 
direction. I therefore venture to bring forward the results 
of an investigation of this subject, on which I have been 
engaged at intervals for the last eighteen months. 

As a first step it appeared desirable to obtain proof, if 
possible, of the particulate nature, or otherwise, of the 
essential cause of vaccinia. 

I next turned my attention to the identification of the 


various micro-organisms which can be grown as the result 
of inoculation of various nutritive media with vaccine 
lymph. And here I should remark that in all the experi- 
ments of this series, calf lymph alone was used, as it 
seemed probable that by these means one would obtain 
more constant results. 

As it was evident that more than one organism could 
usually be obtained by the inoculation of lymph into 
nutrient media, plate cultivations were made, with the 
object of separating out the different varieties present, both 
gelatine and glycerine agar being used for this purpose. 
In this manner I succeeded in obtaining pure growths of 
various micro-organisms, of which those which almost 
invariably occurred included micrococci, apparently much 
resembling, if not identical with, the following : — 

(i) M. Pyogenes aureus; 

(2) M. Cereus flavus ; 

(3) M. Epidermidis ( ? Staphylococcus albus, Klein). 

In these observations I find my experiments are in 
agreement with those of Pfeiffer, while doubtless these 
various micrococci correspond respectively with Buist's 
so-called orange, yellow, and white vaccine. 

As it appeared possible that their exuberant growth 
might be capable of preventing the development of the 
more important organism for which I was seeking, an 
attempt was next made to isolate it by some method of 
treating vaccine lymph,^ which should destroy all extraneous 
micro-organisms without injuring its potency for vaccination. 

Having in mind the method by which Kitasato succeeded 
in obtaining pure cultures of the tetanus bacillus, I con- 


sidered whether it might not be possible, in like manner, 
to isolate the specific organism of vaccinia. For the pur- 
pose of testing the effect of similar measures, I exposed 
capillary tubes of fresh vaccine lymph to different tempera- 
tures for varying lengths of time, afterwards inoculating a 
portion of the contents of these tubes on the calf, a second 
portion being used for the making of plate-cultivations in 
nutrient gelatine and agar-agar. By proceeding in this 
manner, after numerous experiments, I presently arrived at 
a temperature at which those organisms which usually 
grow so luxuriantly when vaccine lymph is inoculated into 
nutrient jelly, are apparently incapable of continued existence 
after exposure to its influence for a certain length of time. 

After a long series of experiments with lymph exposed 
to various temperatures between the limits mentioned, and 
for varying periods of time, I have apparently determined 
that the required temperature is one ranging between 38° 
C. and 42° C. For exposure to the higher of these tem- 
peratures for an hour has the effect of preventing the 
growth in plate-cultivations made subsequently, while at 
the lower temperature a few points of growth are occasion- 
ally seen after the lapse of a day or so. At the same time, 
however, the higher temperature appears occasionally to 
exert an injurious effect on the lymph, as far as regards the 
normal vesiculation which should result from the inoculation 
of vaccine lymph. Further experiments are therefore 
needed before the most suitable temperature and most 
desirable length of exposure can be definitely determined. 

Early in my experiments, my attention was called to the 
fact that lymph stored in capillary tubes nearly always 


becomes cloudy after a longer or shorter interval, while at 
the same time it often becomes uncertain in its action 
when subsequently used for vaccination. 

I endeavoured therefore, in the first place, to determine 
the reason for the opacity which occurs in stored lymph ; 
and, secondly, to find, if possible, some means of preventing 
such an occurrence. As regards the first point, experi- 
ments were carried out as follows : — A large number of 
capillary tubes were filled with calf lymph, every precaution 
as to cleanliness and careful sealing of the tubes being 
observed. These tubes were then set aside, not, as is 
usually the case, lengthwise, but on end. After the lapse 
of a few weeks all the tubes presented little points of 
opacity, and on careful examination it was obvious that 
each of these occurred where a surface of the lymph was in 
contact with a bubble of air. Moreover, it was always at 
the lower end of the line of lymph that such opaque points 
were found, thus showing that they were composed of 
something possessing a higher density than the lymph 
itself, as otherwise there was no apparent reason why they 
should not also be met with at the upper' limit of each 
thread of lymph where air was also present. 

This appearance was quite independent of any coagula- 
tion in the lymph ; indeed, on breaking some of the tubes 
in which it was most marked, no coagulum was found. 
On the other hand, where clotting had taken place after 
the lymph was stored, the opacity was often not in discrete 
points as in the other tubes, but formed with the coagulum 
a central whitish thread in the midst of a clear fluid. In 
these cases, the tiny particles of which an opaque point 
was obviously composed had apparently become more or 
less entangled with the fine thread of fibrin which had 
resulted from the process of coagulation. 


Cultivation experiments were then carried out, gelatine 
tubes being inoculated from opaque points in stored tubes, 
and also, as a control, from tubes of comparatively fresh 
lymph, the capillary tubes used in these experiments 
having been sterilised previous to their being filled with 
lymph. In each case, plate -cultures were made from 
dilutions of the gelatine tubes first inoculated. As a result, 
many more colonies were found in the plates poured from 
inoculations of the old tubes than from those descended 
from the fresher lymph. We are therefore apparently 
justified in considering that the opacity of old-stored lymph 
is, in the main, the outcome of an enormous multiplication 
of aerobic bacteria, the ancestors of which are present in 
the lymph when first taken, although their numbers are 
then so comparatively small as not to render it in any way 

As I have already shown, a certain amount of heat, 
when applied to lymph in sealed capillary tubes, inhibits 
the growth of these aerobic bacteria, while a still higher 
temperature kills them. The exposure of freshly -stored 
lymph, then, to a proper temperature for a sufficient length 
of time, ought to prevent the subsequent appearance of 
opacity, and such is apparently the case. Without great 
care, however, there is in this method considerable danger 
of rendering the lymph inert. 

A simpler method of obtaining the desired result, how- 
ever, is found in the admixture of the lymph with a certain 
proportion of 50 per cent glycerine in distilled and sterilised 
water prior to storage in capillary tubes, which also should 
have previously been sterilised by heat. 

(Miiller showed, long ago, that lymph might be diluted 
with three times its bulk of such a mixture and still retain 
its properties unimpaired, a fact which has been taken 



advantage of by more than one purveyor of trade lymph. 
Experiment shows that not only is this so, at any rate for 
a considerable time, but also that, in tubes filled with such 
diluted lymph, opacity does not apparently result. The 
glycerine appears to inhibit the growth of those aerobic 
bacteria, which in former parts of this paper I have termed 
''extraneous," signifying by that term that their presence 
is not in any way essential, indeed, probably rather the 
reverse, to the successful action of the vaccine lymph. . . . 
Another argument, in favour of the use of lymph diluted 
in this way with glycerine, is found in the fact that, as 
before stated, such a mixture does not dry up nearly as 
readily as ordinary lymph, and therefore in the hands of 
operators, all but the most expert, affords greater facilities 
towards the attainment of a uniformly successful series of 
vaccinations. The fact also that a much larger supply of 
lymph would thus be available is so obvious that it needs 
not to be insisted on. 


Academic de Medecine ; animal 
vaccination in Fiance, 144- 


Adams ; inoculation, 25 

, , variolation of bovine ani- 
mals, 47 

Admixture of glycerine with 
vaccine lymph ; Cheyne, 155- 
156 ; no new device, 155-158 

Aerobic bacteria cause of opacity 
in vaccine lymph, 83-84 

Aitken ; smallpox, 17 

Allen ; inoculation, 21-23 

Amsterdam, animal vaccination 
introduced to, 146 

Animal vaccination, 142-152: 
Ballard on introduction of, 
142-143; Berlin, 211-218; 
Brussels, 147-148, 206-211 ; 
Chambon, 143-145 ; Cologne, 
224-232 ; definition of, 142 ; 
Dresden, 218-224 ; effect of 
on erysipelas and tuberculosis, 
151-152 ; England, 148-152 ; 
English Royal Commission, 
149 ; France, Royal Com- 
mission, 144 - 145 ; Galbiati, 
150 ; Geneva, 232-236 ; Ger- 
many, 147 ; grounds on which 
advocated, 149-152; Holland, 
146 ; introduced to Amster- 
dam, 146 ; introduced to Berlin 
by Pissin, 145 - 146 ; intro- 
duced to Brussels by Warlo- 

mont, 145 ; introduced to 
Paris by Lanoix, 143 - 144 ; 
introduced to Rotterdam, 146 ; 
introduced to Switzerland by 
Haccius, 146 ; introduced to 
Vienna, 146 ; introduction of, 
Ballard and Negri, 142-143 ; 
Paris, 143-144, 146-147, 196- 

Animal Vaccine Estabhshment, 
Government, source of lymph 
used at, 71 ; vaccinations from 
a single calf at, 192 ; vaccina- 
tion at, 106 

Anthony; micro-organisms in 
vaccine lymph, 96-97, 99 

Antitoxin, 134-141 

Arloing : experiments with horse- 
pox lymph, 101-102 ; specific 
micro - organism of vaccine 
lymph, 102 

Arm - to - arm vaccination : and 
erysipelas, 151-152 ; and tuber- 
culosis, 151- 152 ; deterioration 
of, 149-150; Jenner's first 
attempt to carry on, 67-68 ; in 
England, 149 ; English Royal 
Commission on risks incurred 
from, 149-150 ; syphilis con- 
veyed by, 151 ; Woodville, 

Athens ; inoculation in, 20 

Auch6 ; treatment of smallpox by 
serum, 140 



Austria, quality of glycerine in, 

Babes ; micrococci isolated from 
vaccine lymph, 90 

Bacilli : diphtheria, in lymph, killed 
by addition of glycerine, 161 ; 
in calf and variolous lymphs, 
Klein, 98 ; in vaccine lymph, 
Anthony, 96-97, 99, Copeman, 
100, and Kent, 100 

Bacillus, tubercle, action of gly- 
cerine on, Blaxall and Copeman, 
162, 165-168, and Klein, 166- 
167; tubercle, in vaccine lymph, 

Bacteria : aerobic, cause of opacity 
in vaccine lymph, 83-84 ; in- 
fluence of glycerine on growth 
of, Blaxall and Copeman, 162- 
168 ; in vaccine lymph. Crook- 
shank, 95-96 ; specific, of 
vaccine lymph, Koch, 90-91 

Bacteriological examination of gly- 
cerinated calf lymph, 184-185 ; 
purification and preservation of 
vaccine lymph by glycerine, 153 

Bacteriology, 85-127 ; of vaccine 
lymph in 1891, Copeman, 

Badcock ; variolation of bovine 
animals, 47, 60 

Baillard ; effect of glycerine on 
vaccine lymph, 99 ; micro- 
organisms in vaccine lymph, 99 

Ballard ; introduction of animal 
vaccination, 142-143 

Bamberand King ; action of lano- 
line and vaseline on vaccine 
lymph, 173-174 

Baragi ; specific micro-organism 
of vaccine lymph, 91-92 

Baron ; smallpox, 7 

Beale, Lionel ; granules in vaccine 
lymph, 85-86 

Beaugency, natural cow-pox dis- 
covered at, 144 

B^clere ; treatment of smallpox by 
serum, 140 

Berlin : animal vaccination intro- 
duced to, by Pissin, 145-146 ; 
Commission, micro - organisms 
in vaccine lymph, 108, 115 ; 
lymph, results of vaccination 
with, Cory, 217-218 ; vaccina- 
tion arrangements at, 211- 

Berthel ; variolation of bovine 
animals, 50 

Besser ; sp>ecific micro-organism 
of variola, 96 

Beumer ; protective power of 
serum from vaccinated calves, 

Blaxall : action of lanoline and 
vaseline on vaccine lymph, 173- 
180 ; micro - organisms in 
vaccine lymph, 104-108, 114- 
117 ; purification and preserva- 
tion of vaccine lymph by glycer- 
ine, 173-181 

Blaxall and Copeman : action of 
glycerine ; on certain yeasts, 
164-165; on growth of bacteria, 
162-168 ; on tubercle bacillus, 

Boureau ; micro - organisms in, 
and potency of vaccine lymph, 

Bovine animals : and vaccination, 
111-114; inoculation of, with 
variola, 41-66, 71-72 ; sub- 
cutaneous injection of vaccine 
lymph in, Chauveau, 136 

Boyce ; protective power of serum 
from vaccinated calves, 138 

Bruce Low ; increase of vaccine 
material by addition of glycer- 
ine, 157 

Brussels, animal vaccination in, 
145, 147-148, 206-211 

Bryce ; cow-pox in the cow, 

Buchanan, Sir George : animal 



vaccination in England, 148 ; 
dilution of lymph with glycerine, 
171 ; Riigen outbreak of dis- 
ease, I 70-171 
Buist ; micro-organisms in vac- 
cine and variolous lymphs, 92- 

Burdon-Sanderson : granules in 

vaccine lymph, 88 
Buttersack ; granules in vaccine 

lymph, 99-100 

Calf lymph : bacillus in, Klein, 
98 ; English Royal Commis- 
sion on, 168-170 ; viscidity of, 

Calf lymph, glycerinated : advan- 
tages of, Copeman, 1 81-182 ; 
bacteriological examination of, 
184-185 ; collection of lymph 
pulp, 183-184; duration of 
activity of, 185-186 ; effect of, 
on tubercle, 152 ; English 
Royal Commission, 169-170 ; 
inoculation of the calf, 183 ; 
preparation of, 182-187 ; pre- 
paration of the calf, 182-183 ! 
quantity of lymph pulp ob - 
tained, 184 ; treatment of 
lymph pulp, 184 ; use of, de- 
cided on, in England, 149 ; 
reasons for use of, 168-169 ; 
vaccination with, desirable, 

Calf : skin of, changes produced 
in by vaccination, Gustav 
Mann, 74-81 ; to-arm vaccina- 
tion at Nanterre, 202-203 ; 
vaccination, action of leuco- 
cytes in, 77-80, 82 ; vaccina- 
tion, most characteristic feature 
of, 80-81 

Calves : inoculated with smallpox 
[see Variolation) ; vaccinated, 
protective power of serum from, 
Bdclere, 138-140, Beumer, 
138, Boyce, 138, Chambon, 

138-140, Hlava, 138, Houl, 
138, Kinyoun, 138, Kramer, 
138, Menard, 138-140, Peiper, 

Carmichael ; micro-organisms in 
vaccine lymph, 93 

Casual cow-pox, 36-38 

Catherine of Russia inoculated 
by Dimsdale, 23-24 

Causes and effects of variolas 
vaccinae, Jenner, i 

Ceely : cow-pox in the cow, 36 ; 
lymph stocks, 71 ; variolation 
of bovine animals, 44-47, 57, 

Chalybaus ; lymph - triturating 
machine, description of, 222- 

Chambon : action of glycerine on 
vaccine lymph, 158-159 ; ani- 
mal vaccination, 143-145 ; 
micrococci in vaccine lymph, 
96 ; natural cow-pox at St. 
Mand6, 144-145 ; protective 
power of serum from vaccinated 
calves, 138-140 

Chaumier ; micro-organisms in, 
and potency of vaccine lymph, 

Chauveau : contagium particulate, 
proof of, 88-89 ; cow-pox and 
horse - pox, 35 ; dilution of 
vaccine lymph, 88-89 I genera- 
lised vaccinia in solipeds, 116 ; 
granules in vaccine lymph, 86- 
88 ; subcutaneous injection of 
vaccine lymph in bovines and 
horses, 136 ; variolation of 
bovine animals, 48-50, 61-62, 

64. 13s 
Chemistry and morphology of 

vaccine lymph, 82-84 
Chester, smallpox epidemic, 

1774 ; Haygarth, 3-4 
Cheyne ; admixture of glycerine 

with vaccine lymph, 155-156 
China, artificial communication 



of smallpox by respiratory tract 
in, 19 

China, smallpox of great antiquity 
in, S 

Clarke, Jackson : protozoa, 121 

Clarke ; lymph obtained from, by 
Jenner, 69 

Cohn : morphology of vaccine 
lymph, 89-90; opacity of stored 
vaccine lymph, 90 

Collins : cow-pox and smallpox, 
65; the variolous test, 132-133 

Cologne : vaccination arrange- 
ments at, 224-232 ; lymph, 
results of vaccination with, 
Cory, 228 

Commission, English Royal : 
animal vaccination, 149 ; calf 
lymph, 168-170 ; glycerinated 
calf lymph, 169-170 ; quality 
of vaccine lymph, 168-169 1 
risks incurred from arm-to-arm 
vaccination, 150-151 

Commission, German : action of 
glycerine on vaccine lymph, 
160- 16 1 ; extent of lymph 
dilution by glycerine, 161 

Congress, International, of Hy- 
giene, London ; glycerinated 
lymph, 153 

Constantinople, first vaccination 
performed at, 70 

Contagium particulate, proof of ; 
Chauyeau, 88-89 

Continent, etc., source of lymph 
first used on, 69 

Copeman : advantages of glycerin- 
ated calf lymph, 181- 182; 
bacteriology of vaccine lymph 
in 1 89 1, 237-242 ; cultivation 
of variola vaccine, 109-114 ; 
glycerinated lymph, 153-187 ; 
micro - organisms in vaccine 
lymph, 95, 100, 104- 117; 
Riigen outbreak of disease, 
171-172 ; treatment of small- 
pox by serum, 141 ; variolation 

of bovine animals, 58-61, 63- 


Copeman and Blaxall : action of 
glycerine ; on certain yeasts, 
164-165; on growth of bacteria, 
162-168 ; on tubercle bacillus, 

Copeman and Sir Richard 
Thorne ; foreign vaccination 
arrangements, 196-236 

Cornil ; micrococci isolated from 
vaccine lymph, 90 

Corpuscles, red, in vaccine 
lymph, Husband, 82 

Cory : animal vaccination in 
England, 148 ; results of vac- 
cination with Berlin lymph, 
217-218 ; with Cologne lymph, 
228 ; with Dresden lymph, 
221 ; with Paris lymph, 205 ; 
variolation of bovine animals, 
50-51, 56-57 

Cow - pox : and horse - pox, 
Chauveau, 35 ; and smallpox 
identical, controversy as to, 
42 ; casual, 36-38 ; Collins, 65 ; 
derived from horse-pox, 32-34 ; 
Hunter, 29 ; in the cow, Ceely 
and Bryce, 34-36 ; inoculated 
in man, 38-40; Jenner, 28- 
29, 31-32, 43-44 ; natural, dis- 
covered at Beaugency and St. 
Mandd, 144 ; Negri, 143 ; 
Picton, 65 ; references, 40 ; 
varioloe vaccince known as, i 

Cows inoculated with smallpox 
{see Variolation) 

Cozette ; micro - organisms in 
vaccine lymph, 103 

Creighton ; smallpox, 13-14 

Crookshank ; bacteria in vaccine 
lymph, 95-96 

Crusades, spread of smallpox 
during, 12 

Crust, vaccine, description of, 74 

Cultivation of variola vaccine ; 
Copeman, 108-114 



Deaths from smallpox : Iceland, 
1 707- 1 709, 5 ; large proportion 
contributed by children under 
ten years, 4 ; London, 1694- 
1719, 16-17; McVail, 4-5; 
Norwich, 17 10, 17 

De Carro ; lymph obtained from 
Lombardy, 69-70 

Delobel ; micro-organisms in vac- 
cine lymph, 103 

Depaul ; animal vaccination in 
France, 144-145 

Dilution of vaccine lymph : Chau- 
veau, 88-89 ; Reiter, 89; 
Schultz, 89 

Dimsdale : inoculation, 26 ; in- 
oculation of Catherine of 
Russia, 23-24 

Diphtheria bacilli in lymph killed 
by addition of glycerine, 161 

Doering's lymph - grinding ma- 
chine, description of, 213-215 

Domiciliary vaccination, Paris, 

Dresden : lymph, results of vac- 
cination with ; Cory, 221 ; 
vaccination arrangements at, 

Edinburgh: smallpox, 1764- 

1783. 4 

England : admixture of glycerine 
with lymph patented by Warlo- 
mont in, 158; animal vaccina- 
tion in, 148 - 152 ; arm- 
to -arm vaccination in, 149 ; 
quality of glycerine in, 172-173; 
smallpox first introduced into, 
13 ; smallpox in seventeenth 
century in, 15-17 ; use of calf 
lymph decided on, in, 149 

English Royal Commission : 
animal vaccination, 149 ; calf 
lymph, 168-170 ; glycerinated 
calf lymph, 169-170; risks 
incurred from arm - to - arm 
vaccination, 1 50-1 51 ; variola- 

tion of bovine animals, 60- 

Epidemic smallpox : Chester, 
1774 ; Haygarth, 3 ; Iceland, 
1707-1709, 5 ; Ware, 1722, 3 ; 
Warrington, 1773, 3 

Epidemics, Kilmarnock, 1728- 
1763, 4 ; McVail, 5 

Equine origin of Lombardy lymph, 

Erysipelas : and arm-to-arm vac- 
cination, 151 - 152 ; effect of 
animal vaccination on, 151-152 ; 
streptococcus of, in vaccine 
lymph, Klein, 95 

Esthn ; distribution of lymph, 71 

Eternod ; variolation of bovine 
animals, 55, 60 

Europe, smallpox unknown in 
prior to end of sixth century, 5 

"Extraneous" organisms: in- 
fluence of glycerine upon the 
growth of, 153-182; in vaccine 
lymph, 104-108 ; removal of, 
by heat not advantageous, 154- 

Fee for inoculation, 27-28 

Feiler ; specific microbe in vaccine 
lymph, 91-92 

Ferroni ; protozoa, 122 

Fischer ; variolation of bovine 
animals, 52-54 

Foreign vaccination arrangements ; 
Sir Richard Thorne and Cope- 
man, 189-236 

Foster, Michael ; efficiency of 
lymph stocks, 128 

France : animal vaccination in ; 
Acad^mie de M^decine, 144- 
145 ; smallpox in, 6 

Freyer : specific organism of vac- 
cine lymph, 115-116 ; vaccina- 
tion by spleen juice, etc., 115- 

Galbiati : propagation of other 



human diseases with vaccinia, 
150 ; retro-vaccination, 143, 

Gassner ; variolation of bovine 

animals, 42-43 
Garr6 ; micro-organisms in vaccine 

lymph, 94 
Generalised vaccinia in solipeds ; 

Chauveau, 116 
Geneva, vaccination arrangements 

at, 232-236 
German Commission : action of 

glycerine on vaccine lymph, 

160 - 161 ; extent of lymph 

dilution by glycerine, 161 
Germany, animal vaccination in, 


Glycerinated calf lymph : advan- 
tages of; Copeman, 181 -182; 
and International Congress of 
Hygiene, London, 153 ; bac- 
teriological examination of, 184- 
185 ; collection of lymph pulp, 
183-184; Copeman's investiga- 
tions, 153-187; duration of 
activity of, 185-186 ; effect of 
on tubercle, 152 ; English 
Royal Commission on, 169- 
170 ; inoculation of the calf, 
183 ; preparation of, 182-187; 
preparation of the calf, 182- 
183 ; quantity of lymph pulp 
obtained, 184 ; treatment of 
lymph pulp, 184 ; use of, de- 
cided on, in England, 149 

Glycerinated vaccine lymph, dilu- 
tion of, Schultz, 89 

Glycerine, action of : on certain 
yeasts, Blaxall and Copeman, 
164-165 ; on growth of bacteria, 
Blaxall and Copeman, 162-168 ; 
on " extraneous " micro-organ- 
isms, 153-182 ; on tubercle 
bacillus, Blaxall and Copeman, 
165-168, and Klein, 166-167 ; 
on Vaccine Lymph, Baillard, 
99, Chambon, 158-159, Frosch, 

160-161, German Commission, 
160-161, Kitasato, 161, Klein, 
160 ; Koch, 160- 161 ; Leoni, 
99, 159-160; Menard, 103, 
158-159 ; Pfeiffer, 160-161 ; 
Schmidtmann,i6o-i6i ; Strays, 
159 ; admixture of with vaccine 
l3^mph, Cheyne, 155-156; ad- 
mixture of with vaccine lymph, 
no new device, 155-158 ; ad- 
mixture of with vaccine lymph, 
patented in England, Warlo- 
mont, 158 ; analyses of, Wilson 
Hake, 172-173 ; and thymol, 
lymph mixed with, cause of dis- 
ease in Riigen, 170-171 ; diph- 
theria bacilli in lymph killed by 
addition of, 161 ; germicidal 
value of, 161-162; increase of 
vaccine material by addition of, 
Bruce Low, 1 57, Mackenzie, 1 57, 
and Muller, 156-157 ; potency of 
lymph impaired by chemical 
agents other than, 161 ; 
purification and preservation 
of vaccine lymph by, 153, 
173-18 1 ; purity of, past and 
present, 171-172 ; quality of, in 
Austria and in England, 172- 
173; Sir George Buchanan on 
dilution of lymph with, 171 ; 
Startin's advocacy of thera- 
peutic applications of, 156 

Gourny ; micro - organisms in 
vaccine lymph, 103 

Government lymph laboratories, 

Granules in vaccine lymph ; Beale, 
Lionel, 85-86, Burd on -Sander- 
son, 88, Buttersack, 99-100, 
Chauveau, 86-88, Hallier, 86, 
Keber, 86, Sacco, 85, Zurn, 

"Grease," or horse-pox, 31-35 

Gregory ; inoculation, 26 

Grigorieu ; micro - organisms in 
vaccine lymph, 94 



Guttman ; micro - organisms in 

vaccine lymph, 92 
Guarneri ; protozoa, 118-119 
Guarneri's "parasites," 'j'j, 119- 

123, 125-126 

Haccius : animal vaccination in- 
troduced into Switzerland by, 
146 ;• variolation of bovine ani- 
mals, 55-56, 60 

Hallier ; granules in vaccine 
lymph, 86 

Haygarth ; smallpox epidemic, 
Chester 1774, 3 

Hervieux ; variolation of bovine 
animals, 63-64 

Hime ; variolation of bovine ani- 
mals, 54, 60 

Histology of the vaccine vesicle, 


Hlava: micro-organisms in vaccine 
lymph, 93-94 ; protective power 
of serum from vaccinated calves, 
138 ; protozoa, 117 

Holland, animal vaccination in, 

Horse-pox: and cow-pox, Chau- 
veau, 35 ; cow - pox derived 
from, 32-35 ; experiments as 
to. Tanner, 32 ; Loy, 32 ; 
lymph, experiments with, Arlo- 
ing, 101-102; Sacco, 32-34, 70 

Horses, subcutaneous injection of 
vaccine lymph in ; Chauveau, 

Houl ; protective power of serum 
from vaccinated calves, 138 

Human diseases, other, propaga- 
tion of, with vaccinia, 150-152 

Hunter ; cow-pox, 29 

Husband ; red corpuscles in vac- 
cine lymph, 82 

Hygiene, International Congress 
of, and glycerinated lymph, 153 

Iceland, smallpox deaths, 1707- 
1709. 5 

India, lymph conveyed to, 70 ; 
smallpox of great antiquity in, 5 

Inoculated cow-pox in man, 38-40 

Inoculation, 19-28, 67 : Adams, 
25; Allen, 21-23; brothers 
Sutton, 24-26 ; controversy as 
to efficiency of, 25-27 ; diffusion 
of ; Adams, Dimsdale, and 
brothers Sutton, 23 ; Dimsdale, 
26 ; fatahty from, 26-27 I ^^^ 
for, 27-28 ; first introduced into 
England by Lady Montagu, 
i8, 20-21 ; Gregory, 26 ; in 
Athens and Smyrna, 20 ; Eng- 
land, first operation ; Maitland, 
21 ; of condemned felons, 21 ; 
George I., 21 ; origin of, un- 
known, 19 ; practised in Turkey, 
19 ; prohibited since 1840, 18; 
Sir Thomas Watson, 26-27 

Italy, smallpox in, 6 

Jenner : cow-pox, 28-32, 43-44 ; 
deterioration of arm - to - arm 
vaccination, 149 - 150 ; full 
degree of activity of vaccine 
lymph, 150 ; his first attempt 
at arm-to-arm vaccination, 67- 
68 ; his first vaccination, 29-30, 
67 ; proved vaccination to be a 
protection from smallpox, 30 ; 
vaccinia, 41-42 ; variolae vac- 
cinae, I 

Juhel ; variolation of bovine 
animals, 63 

Keber ; granules in vaccine lymph, 

Kent ; bacilli in vaccine lymph, 

100 ; vaccine vesicle, 74 
Kilmarnock, smallpox, 1728- 

1763. 4 

King and Bamber : action of 
lanoline and vaseline on vaccine 
lymph, 173-174 ; variolation of 
bovine animals, 53-54, 60 

Kinyoun : protective power of 



serum from vaccinated calves, 
138 ; treatment of smallpox by 
serum, 141 

Kitasato ; action of glycerine on 
vaccine lymph, 161 

Klebs ; micro - organisms in 
vaccine lymph, 90 

Klein : action of glycerine on 
tubercle bacillus, 166-167 ; and 
on vaccine lymph, 160 ; bacilli 
in calf and variolous lymphs, 
98-99 ; streptococcus of ery- 
sipelas found in vaccine lymph, 
95 ; variolation of bovine 
animals, 56-58, 61, 63, 135 

Koch: micro-organisms in vaccine 
lymph, 90-91 ; rinderpest, 44 ; 
specific bacteria of vaccine 
lymph, 90-91 

Kramer; protective power of serum 
from vaccinated calves, 138 

Laboratories, Government Lymph, 

Landmann : micro-organisms in 
vaccine lymph, loo-ioi ; treat- 
ment of smallpox by serum, 

Lanoix ; animal vaccination intro- 
duced to Paris by, 143-144 ; 
natural cow-pox at St. Mand6 
discovered by, 144 

Lanoline, action of, on vaccine 
lymph; Blaxall, 173-180 ; King 
and Bamber, 173-174 

Layet ; variolation of bovine 
animals, 63-64 

Le Dantec : micro-organisms in 
vaccine lymph, loi ; variola- 
tion of bovine animals, 64 

Leoni : action of glycerine on 
vaccine lymph, 99, 159-160 ; 
micro - organisms in vaccine 
lymph, 95 

Leucocytes, action of, in calf 
vaccination, 77-80, 82 

London, 1694- 171 9, deaths from 

smallpox in, 16-17 ; smallpox 
epidemics, McVail, 5 

Loy ; horse-pox, 32 

Lymph: Berlin, results of vaccina- 
tion with, Cory, 217-218 ; calf, 
viscidity of, 82 ; Cologne, 
results of vaccination with, 
Cory, 228 ; dilution of, with 
glycerine, extent of, German 
Commission, 161, andSirGeorge 
Buchanan, 171 ; diphtheria 
bacilli in, killed by addition of 
glycerine, 161 ; Dresden, results 
of vaccination with, Cory, 221 ; 
glycerinated,Copeman's investi- 
gations, 153-187 ; granules in, 
Beale, Lionel, 85-86; Burdon- 
Sanderson, 88 ; Buttersack, 99- 
100; Chauveau, 86-88; Hallier, 
86 ; Keber, 86 ; Sacco, 85 ; 
Zurn, 86 ; International Con- 
gress of Hygiene, London, 153 ; 
grinding - machine, Doering's, 
description of, 213-215 ; leuco- 
cytes present in, 82 ; Lombardy, 
of equine origin, 70 ; mixed 
with glycerine and thymol cause 
of disease in Riigen, 170-171 ; 
obtained from Clarke by Jenner, 
69 ; Lombardy, Sacco, and De 
Carro, 69-70 ; micro-organisms 
in, 82-83; Anthony, 96-97, 
99 ; Babes, 90 ; Baillard, 99 ; 
Berlin Commission, 108, 115 ; 
Blaxall, 104-108, 114- 117; 
Boureau, 102 ; Buist, 92-93 ; 
Carmichael, 93; Chambon, 96; 
Chavunier, 102 ; Copeman, 95, 
100, 104-117 ; Cornil, 90 ; Co- 
zette, 103; Crookshank, 95-96; 
Delobel, 103 ; Garr^, 94 ; 
Gourny, 103 ; Grigorieu, 94 ; 
Guttman, 92 ; Hlava, 93-94 ; 
Kent, loo ; Klebs, 90 ; Klein, 
98-99, 106; Koch, 90-91; 
Landmann, 100 - loi ; Le 
Dantec, loi ; Leoni, 95 ; Mai- 



jean, 97; Marotla, 92; Mai'tin, 
loi ; Meguin, 92 ; Menard, 
96, 102-103 ; Paul, 103-104 ; 
Pfeiffer, L. , 94 ; Protopopoff, 
95 ; Quist, 91 ; Ruete, 100 ; 
Sacqu^p^e, 103 ; Straus, 96 ; 
Tenholt, 93 ; Woitow, 94-95 ; 
microscopical examination of, 
82 ; morphology of, Cohn, 89- 
90 ; Paris, results of vaccination 
with, Cory, 205 ; Passy, origin 
of, 70-71 ; potency of, Boureau 
and Chaumier, 102 ; potency 
of, impaired by chemical agents 
other than of glycerine, 161 ; 
potency of, unimpaired by addi- 
tion of glycerine, Miiller, 156- 
157 ; source of, first used on 
Continent, etc. , 69 ; source of, 
used at Government Animal 
Vaccine Establishment, 71 ; 
specific micro - organism of, 
Arloing, 102 ; Baragi, 91-92 ; 
Feiler, 91-92; Freyer, 115-116 ; 
Koch, 90-91 ; Quist, 91 ; Sere- 
briakoff, 91-92 ; Voigt, 92 ; 
sterilised, experiments with, 
134-135; stocks, Ceely, 71; 
stocks, efficiency of, Michael 
Foster, 128 ; stocks, various, 
historyof, 67-72; stored, opacity 
of, Cohn, 90 ; streptococcus of 
erysipelas found in, Klein, 95 ; 
subcutaneous injection of, in 
bovines, horses, and monkeys, 
136; triturating machine, Chaly- 
baus, description of, 222-224 ; 
tubercle bacillus in, 108 ; used 
in vaccinal and variolous tests 
in the monkey, 131 ; variola 
vaccine, 71 ; variolous, micro- 
organisms in, Buist, 93 ; Wood- 
ville's, 68 

Lymphs, calf and variolous, bacilli 
in, Klein, 98 

Lyons Commission ; variolation of 
bovine animals, 48-49, 6i 

Macaulay ; smallpox, 2 

MacEUiot ; treatment of smallpox 
by serum, 141 

Mackenzie ; increase of vaccine 
material by addition of gly- 
cerine, 157 

McVail ; deaths from smallpox, 


Maitland ; first to inoculate in 
England, 21 

Maljean ; micrococci in vaccine 
lymph, 97 

Mand^, St. ; natural cow-pox dis- 
covered at, 144 

Mann, Gustav; changes produced 
in skin of calf by vaccination, 

Marotla ; micro - organisms in 
vaccine lymph, 92 

Martin ; micro - organisms in 
vaccine lymph, loi ; variolation 
of bovine animals, 49, 61, 64 

Massari ; protozoa, 122 

Measles, smallpox confounded 
with, 8 

Meguin ; micro - organisms in 
vaccine lymph, 92 

Menard : action of glycerine on 
vaccine lymph, 103, 158-159 ; 
micro - organisms in vaccine 
lymph, 96, 102-103 ; protective 
power of serum from vaccinated 
calves, 138-140 

Micro-organisms : " extraneous," 
influence of glycerine upon the 
growth of, 153-182 ; removal 
of, by heat not advantageous, 
154-155 ; in vaccine lymph, 
82-83 ; Anthony, 96-97, 99 ; 
Babes, 90 ; Baillard, 99 ; Berlin 
Commission, 108, 115; Blaxall, 
104-108, 114-117; Boureau, 
102 ; Buist, 92-93 ; Carmichael, 
93 ; Chambon, 96 ; Chaumier, 
102 ; Copeman, 95, 100, 104- 
117; Cornil, 90; Cozette, 103 ; 
Crookshank, 95-96 ; Delobel, 



103 ; Garr6, 94 ; Gourny, 103 ; 
Grigorieu, 94 ; Guttman, 92 ; 
Hlava, 93-94 ; Kent, 100 ; 
Klebs, 90 ; Klein, 98-99, 106 ; 
Koch, 90-91 ; Landmann, 100- 
loi ; Le Dantec, loi ; Leoni, 
95 ; Maljean, 97 ; Marotla, 
92 ; Martin, loi ; Meguin, 
92 ; Menard, 93, 96, 102-103 I 
Paul, 103-104 ; Pfeiffer, L. , 94 ; 
Protopopoff, 95 ; Quist, 91 ; 
Ruete, 100 ; Sacqu^pte, 103 ; 
Straus, 96 ; Tenholt, 93 ; 
Woitow, 94 - 95 ; in variolous 
lymph, Buist, 93 ; specific, of 
vaccine lymph, Baragi, 91-92 ; 
Feiler, 91-92; Freyer, 115- 
116 ; Koch, 90-91 ; Quist, 
91 ; Serebriakoff, 91 - 92 ; 
Voigt, 92 

Monkey : complete vaccination 
and variolation in the, proved, 
130-132 ; course of vaccinia 
and variola in the, 129-130 ; 
difference between vaccinia and 
variola in the, 130 ; lymph 
used in vaccinal and variolous 
tests in the, 131 ; method of 
vaccination in the, 128-129; 
subcutaneous injection of ; 
vaccine lymph in the, 136 ; 
susceptibility of, to vaccinia and 
variola, 128-130 ; variola and 
vaccinia in the, 128-133 I vario- 
lated, blood plasma of, as pro- 
tection against vaccination, 136- 

Montagu, Lady, inoculation first 
introduced into England by, 
18, 20-21 

Monti; protozoa, 119 

Moore, Norman ; smallpox, 14 

Morphology and chemistry of 
vaccine lymph, 82-84 ; mor- 
phology of vaccine lymph, 
Cohn, 89-90 

Miiller : potency of lymph un- 

impaired by addition of gly- 
cerine, 156-157 ; protozoa, 124 

Nanterre, calf-to-arm vaccination 
at, 202-203 

Negri^: introduction of animal 
vaccination, 142-143 ; natural 
cow-pox, 143 ; retro-vaccina- 
tion, 143 

Norwich, 1710, deaths from 
smallpox in, 17 

Ogata ; protozoa, 121 

Opacity in vaccine lymph, causes 

of, 83 ; in stored vaccine lymph, 

Cohn, 90 

Papular stage of vaccination, 73 
" Parasites," Guarnieri's, tj, 119- 

123, 125-126 
Paris : animal vaccination at, 146- 
147, 196-206 ; animal vaccina- 
tion introduced to, Lanoix, 
143-144 ; crude calf lymph in 
use at, 190-191 ; description 
of Institut de Vaccine Animale 
at, 196-201 ; domiciliary vac- 
cination at, 203-206 ; lymph, re- 
sults of vaccination with, Cory, 
205 ; street vaccination at, 204- 
205; vaccine station of Acad(Jmie 
de M<5decine at, 201-202 
Passy lymph, origin of, 70-71 
Pathology of vaccinia, 31-40 
Paul ; micro-organisms in vaccine 

lymph, 103-104 
Pearson ; lymph distributed by, 

Peiper ; protective power of serum 

from vaccinated calves, 138 
Pepys ; smallpox, 3 
Pfeiffer, E. ; protozoa, 121 
Pfeiffer, L. ; micro-organisms in 
vaccine lymph, 94 ; protozoa, 
118, 120-121 
Picton : cow-pox and smallpox, 
65 ; the variolous test, 132 133 



Pissiii, animal vaccination intro- 
duced to Berlin by, 145-146 

Plague, comparison with small- 
pox ; Macaulay, 2 

Plasma, blood, of variolated 
monkey as protection against 
vaccination, 136-138 

Plimmer ; protozoa, 119-120 

Potency of lymph ; Mtiller, 156- 
157; BoureauandChaumier,io2 

Propagation of other human 
diseases with vaccinia, 150-152 

Protopopoff; micro-organisms in 
vaccine lymph, 95 

Protozoa, 1 17-126 : Clarke, Jack- 
son, 121 ; Ferroni, 122 ; 
Guarnieri, 118- 119 ; Hlava, 
117; Massari, 122; Monti, 
119 ; Miiller, 124 ; Ogata, 
121 ; Pfeiffer, E. , 121; Pfeiffer, 
L., 118, 120-121 ; PHmmer, 
1 1 9- 1 20 ; Reed, Walter, 123- 

124 ; Renault, 118 ; Rieck, 
118 ; Ruffer, 1 19-120 ; Salmon, 
Paul, 122-123 ; Stokes, 124- 

125 ; Van der Loeff, 121 ; Von 
Sicherer, 121 ; Wegefarth, 124 

Pustular stage of vaccination, 74 
Putnam ; variolation of bovine 
animals, 47 

Quist's experiments ; vaccine 
lymph, 91 

Reed, Walter ; protozoa, 123-124 
References : animal vaccination, 
152 ; antitoxin, 141 ; bacteri- 
ology, 126-127 ; cow-pox, 40 ; 
histology of smallpox and 
vaccine vesicles, 84 ; lymph 
stocks, 72 ; preservation, stor- 
age, and use of lymph, 188 ; 
protozoa, 127 ; relationship of 
variola and vaccinia, 66 ; 
variola and vaccinia, 133 
Reiter ; dilution of vaccine lymph, 

Relationship of variola and 
vaccinia, 41-66 

Renault ; protozoa, 118 

Rhazes ; smallpox, 6, 8-12 

Rieck; protozoa, 118 

Rinderpest ; Koch, 44 

Rotterdam, animal vaccination 
introduced to, 146 

Royal Commission, English : 
animal vaccination, 149 ; calf 
lymph, 168-170 ; glycerinated 
calf lymph, 169-170 ; risks in- 
curred from arm-to-arm vac- 
cination, 1 50-1 5 1 ; variolation 
of bovine animals, 60-61, 65 

Ruete ; micro - organisms in 
vaccine lymph, 100 

Ruffer; protozoa, 119-120 

Rligen, lymph mixed with gly- 
cerine and thymol, cause of 
disease in, 170- 171 ; Cope- 
man, 171-172, and Sir George 
Buchanan, 170- 171 

Sacco : granules in vaccine lymph, 
85 ; horse -pox, 32-34, 70 ; 
lymph obtained from Lom- 
bardy by, 69-70 

Sacquep6e ; micro-organisms in 
vaccine lymph, 103 

Salmon, Paul; protozoa, 122-123 

Schultz ; dilution of glycerinated 
vaccine lymph, 89 

Serebriakoff ; specific micro -or- 
ganism in vaccine lymph, 91-92 

Serum from vaccinated calves, 
protective power of : B^clere, 
138-140; Beumer, 138; Boyce, 
138 ; Chambon, 138 - 140 ; 
Hlava, 138 ; Houl, 138 ; 
Kinyoun, 138 ; Kramer, 138 ; 
Menard, 138-140 ; Peiper, 138 

Serum, treatment of smallpox by : 
Auche, 140 ; Beclere, 140 ; 
Copeman, 141 ; Kinyoun, 
141 ; Landmann, 140 - 141 ; 
MacElliot, 141 



Simpson ; variolation of bovine 
animals, 50-53, 60 

Skin of calf, changes produced 
in, by vaccination ; Gustav 
Mann, 74-81 

Smallpox : Aitken, 17 ; and cow- 
pox identical, controversy as 
to, 42 ; artificial communica- 
tion of, by respiratory tract, 
China, ig ; Baron, 7 ; Chester, 
1774, Haygarth, 3 ; Collins, 
65 ; comparison with plague, 
Macaulay, 2 ; Creighton, 13- 
14 ; death of Queen Mary, 
1694, from, 2 ; deaths, Ice- 
land, 1707- 1 709, 5 ; deaths, 
London, 1694- 1719, 16-17 ; 
deaths, McVail, 4-5 ; deaths, 
none above ten years in War- 
rington 1773 and in Chester 
1774, 4 ; deaths, Norwich 
1710, 17 ; deaths, quarter 
amongst children, 0-12 months, 
Chester 1773, 4 ; during i6th 
century, 15 ; Edinburgh 1764- 
1783, 4 ; England, 17th cen- 
tury, 15-17 ; eruption, stages 
of, 73 ; first introduced into 
England, 13 ; Gaddcsden, John 
of, 13-14 ; Gregory of Tours, 
6 ; history of, 1-18 ; history of 
in three stages, Aitken, 17-18 ; 
houses, first erected, 13 ; India, 
China, Europe, Arabia, Italy, 
France, 5-6 ; inoculation of 
bovine animals with, 41-66 
{^see also Variolation) ; in 17th 
century, 3 ; Kilmarnock 1728- 
1763, 4 ; London, McVail, 5 ; 
Macaulay, 2 ; Marius, Bishop 
of Averche, 6 ; Moore, Nor- 
man, 14 ; Pepys, 3 ; Picton, 
65 ; poets on, 15-16 ; ravages 
of, 2, 12 ; red hangings as 
curative treatment for, 13-14 ; 
Rhazes, 6, 8-12 ; seasons in 
which prevalent, 9 ; spread of 

during Crusades, 12 ; treat- 
ment of, by serum, Auche, 

140, Beclere, 140, Copeman, 

141, Kinyoun, 141, Land- 
mann, 140-141 and MacEUiot, 
141 ; treatment of, Sydenham, 
24 ; Waller, Mrs. , 2-3 ; Ware, 
1722, 3 ; Warrington, 1773, 3 

Smyrna, inoculation in, 20 

Solipeds, generalised vaccinia in ; 
Chauveau, 116 

Spleen juice, etc. , vaccination by, 
Freyer, 115-117 

Startin's advocacy of therapeutic 
applications of glycerine, 156 

Sterilised lymph, experiments 
with, 134-135 

Stokes; protozoa, 124-125 

Straus : action of glycerine on 
vaccine lymph, 159 ; micro- 
organisms in vaccine lymph, 

Subcutaneous injection of vaccine 
lymph in bovines and horses, 
Chauveau, 136 ; in the mon- 
key, 136 

Susceptibility of the monkey to 
vaccinia and variola, 128-130 

Switzerland, animal vaccination 
in, 146 

Sutton brothers ; inoculation, 

Sydenham ; treatment of small- 
pox, 24 

Syphilis conveyed by arm-to-arm 
vaccination, 151 

Tanner ; experiments as to horse- 
pox, 32 

Tenholt ; micro - organisms in 
vaccine lymph, 93 

Thiele, Basil ; variolation of 
bovine animals, 43, 60 

Thome, Sir Richard ; Foreign 
vaccination arrangements, 189- 

Thymol and glycerine, lymph 



mixed with, cause of disease in 
Rugen, 170-17 1 

Troja ; retro- vaccination, 143 

Tubercle bacillus, action of 
glycerine on ; Blaxall and Cope- 
man, 165-168 ; Klein, 166- 

Tubercle bacillus in vaccine 
lymph, 108 

Tuberculosis : and glycerinated 
calf lymph, 151-152 ; and arm- 
to-arm vaccination, 151-152 

Turkey, inoculation practised in, 

Vaccinal and variolous tests in the 
monkey, 130- 131 

Vaccinated calves, protective 
power of serum from : B^clere, 
138-140; Beumer, 138 ; Boyce, 
138; Chambon, 138-140; 
Hlava, 138 ; Houl, 138 ; 
Kinyoun, 138 ; Kramer, 138 ; 
Menard, 138-140 ; Peiper, 138 

Vaccination, 28-40 

Vaccination, animal, 142-152 : 
and erysipelas and tuberculosis, 
151 -152; Berlin, 211 -218; 
Brussels, 147-148, 206-211 ; 
Chambon, 143-145 ; Cologne, 
224-232 ; definition of, 142 ; 
Dresden, 218-224 ! England, 
148-152 ; English Royal Com- 
mission, 149; France, Academic 
de M^decine, 144-145 ; Gal- 
biati, 150 ; Geneva, 232 - 
236 ; Germany, 147 ; grounds 
on which advocated, 149- 
152; Holland, 146; intro- 
duced to Amsterdam, 146, 
Berlin by Pissin, 145-146, 
Brussels by Warlomont, 145, 
Paris, Lanoix, 143-144; Rotter- 
dam, 146, Switzerland by 
Haccius, 146 and Vienna, 146 ; 
introduction of, Ballard and 
Negri, 142-143 ; Paris, 146- 

147, 196-206; at Government 
Animal Vaccine Establishment, 

Vaccination, arm -to -arm : and 
erysipelas, 151 - 152 ; and 
tuberculosis, 151- 152; de- 
terioration of, 149-150 ; in 
England, 149 ; risks incurred 
from, English Royal Com- 
mission, 150-151 ; syphilis 
conveyed by, 151 ; Woodville, 

Vaccination arrangements : Berlin, 
211-218 ; Brussels, 206-211 ; 
Cologne, 224-232 ; Dresden, 
218-224 ; Foreign, Sir Richard 
Thorne, 189-236 ; Foreign, 
Copeman, 196-236 ; Geneva, 
232-236 ; Paris, 196-206 

Vaccinations from a single calf : 
at Government Animal Vaccine 
Establishment, 192 ; when 
lymph is glycerinated, 186, 
192, 216, 227 ; when lymph is 
not glycerinated, 192 

Vaccine crust, description or, 74 

Vaccine lymph : act-ion of glycer- 
ine on, Baillard, 99 ; Chambon 
and Menard, 103, 158-159; 
Frosch, 160- 161; German Com- 
mission, 160- 161 ; Kitasato, 
161 ; Klein, 160 ; Koch, 160- 
161 ; Leoni, 99, 159-160 ; 
Pfeiffer, 160-16 1 ; Schmidt- 
mann, 160 - 161 ; Straus, 
159 ; action of lanoline and 
vaseline on, Blaxall, 173-180; 
King and Bamber, 173-174 ; 
admixture of glycerine with, 
Cheyne, 155-156; no new 
device, 155-158 ; patented in 
England, Warlomont, 158 ; 
appearance of, before maturity, 
82 ; bacteriological purification 
and preservation of, by gly- 
cerine, 153 ; causes of opacity 
in, 83 ; chemistry and mor- 



phology of, 82-84; Copeman, 
in 1 89 1, on bacteriology of, 
237-242 ; dilution of, Chau- 
veau, 88-89, Reiter, 89, 
Schultz, 89, and Sir George 
Buchanan, 171 ; diphtheria 
bacilli in, killed by addition 
of glycerine, 161 ; "extrane- 
ous " micro-organisms in, 104- 
108 ; from equine source, 
32-35. 70 ; full degree of 
activity of, Jenner, 150 ; 
glycerination, extent of, 

German Commission, 161 ; 
lymph, granules in, Beale, 
Lionel, 85 - 86 ; Burdon - 
Sanderson, 88 ; Buttersack, 
99-100 ; Chauveau, 86-88 ; 
Hallier, 86 ; Keber, 86 ; Sacco, 
85 ; Zurn, 86 ; leucocytes 
present in, 82 ; micro-organisms 
in, 82-83 ; Anthony, 96-97, 
99 ; Babes, 90 ; Baillard, 99 ; 
Berlin Commission, 108, 115 ; 
Blaxall, 104-108, 114- 117; 
Boureau, 102 ; Buist, 92-93 ; 
Carniichael, 93 ; Chambon, 96 ; 
Chaumier, 102 ; Copeman, 95, 
100, 104- 117; Cornil, 90; 
Crookshank, 95-96 ; Cozette, 
103 ; Delobel, 103 ; Garr6, 94 ; 
Gourny, 103 ; Grigorieu, 94 ; 
Guttman, 92 ; Hlava, 93-94 ; 
Kent, 100 ; Klebs, 90 ; Klein, 
98-99, 106 ; Koch, 90-91 ; 
Landmann, 100 - loi ; Le 
Dantec, 10 1 ; Leoni, 95 ; 
Maljean, 97 ; Marotla, 92 ; 
Martin, loi ; Meguin, 92 ; 
Mdnard, 93, 96, 102-103; 
Paul, 103-104 ; Pfeiffer, L. , 
94 ; Protopopoff, 95 ; Quist, 
91 ; Ruete, 100 ; Sacqu6p6e, 
103 ; Straus, 96 ; Tenholt, 93 ; 
Woitow, 94-95 ; microscopical 
examination of, 82 ; mixed 
with glycerine and thymol. 

cause of disease in Riigen, 170- 
171 ; morphology of, Cohn, 
89-90 ; potency of, Boureau 
and Chaumier, 102 ; unim - 
paired by addition of glycerine, 
Miiller, 156-157 ; purification 
and preservation by glycerine, 
Blaxall, 1 73- 18 1 ; quality of, 
English Royal Commission, 
168-169 ; Quist's experiments, 

91 ; red corpuscles in. Hus- 
band, 82; specific organism of, 
Arloing, 102 ; Baragi, 91-92 ; 
Feiler, 91-92; Freyer, 115- 
116 ; Koch, 90->9i ; Quist, 91 ; 
Serebriakoff, 91-92, and Voigt, 

92 ; sterilised or filtered, effects 
of, 84 ; sterilised, experiments 
with, 134-135 ; streptococcus 
of erysipelas found in, Klein, 
95 ; stocks, efficiency of, 
Michael Foster, 128 ; stored, 
opacity of, Cohn, 90 ; subcu- 
taneous injection of, in the 
monkey, bovines, and horses, 
Chauveau, 136 ; tubercle bacil- 
lus in, 1 08 ; vesicle, description 
of, 73-74 ; vesicle, histology of, 


Vaccinia and variola : in the 
monkey, 128-133; relation- 
ship of, 41-66 

Vaccinia: generalised, in solipeds, 
Chauveau, 116 ; in the mon- 
key, course of, 129, 130 ; 
Jenner, 41-42 ; pathology of, 
31, 40 ; propagation of other 
human diseases with, 150-152 ; 
specific organism of, Blaxall 
and Copeman, 11 4- 117; sus- 
ceptibility of monkey to, 128- 

Vaccinococcus, description of ; 
Voigt, 92 

Van der Loeff ; protozoa, 121 

Vanselow ; animal vaccination in 
Germany, 147 



Variola : earliest employment and 
derivation of, 7-8 ; specific 
organism of, Besser, 96 ; 
Blaxall and Copeman, 114- 
117 ; susceptibility of monkey 
to, 128-130 ; vaccine, cultiva- 
tion of, Copeman, 108 -114; 
vaccine lymph, 71 

Variolae vaccinae, Jenner, i 

Variolated monkey, blood plasma 
of, as protection against vac- 
cination, 136-138 

Variolation : complete, in the mon- 
key proved, 130-132; of bovine 
animals, 41-66, 71-72 ; Adams 
and Putnam, 47 ; Badcock, 
47, 60 ; Berthel, 50 ; Ceely, 
44-47, 57, 60 ; Chauveau, 
47-50, 61-62, 64, 135 ; Cope- 
man, 58-61, 63-65, 108-114; 
Cory, 50-51, 56-57; Dupuy, 
63 ; English Royal Commis- 
sion, 60-61, 65 ; Eternod, 55, 
60; Fischer, 52-54, 71; Gassner, 
42-43 ; Haccius, 55-56, 60, 71 ; 
Hervieux, 63-64 ; Hime, 54, 
60 ; Juhel, 63 ; King, 53-54. 

60, 71 ; Klein, 56-58, 61, 63, 
135 ; Layet, 63, 64 ; Le 
Dantec, 64 ; Lyons Commis- 
sion, 48-49, 61 ; Martin, 49, 

61, 64; Simpson, 50-53, 60; 
Thiele, Basil, 43, 60 ; Viborg, 
43 ; Voigt, 49-50, 60 

Variola, treatment of, by serum : 
Auch6, 140; B^clfere,i4o; Cope- 
man, 141; Kinyoun,i4i; Land- 
mann, 140-141 ; MacEUiot, 141 

Variolous lymph : bacillus in, 
Klein, 98 ; micrococci in, Buist, 

Variolous test, 68 : Collins and 
Picton, 132-133; in the monkey, 

Vaseline, action of, on vaccine 
lymph: Blaxall, 173-180; King 
and Bamber, 173-174 

Vesicle, vaccine : description of, 
73-74 ; histology of, 73-84 

Viborg ; variolation of bovine 
animals, 43 

Vienna, animal vaccination intro- 
duced to, 146 

Voigt ; vaccinococcus, described 
by, 92 ; variolation of bovine 
animals, 49-50, 60 

Von Sicherer, protozoa, 121 

Waller, Mrs. ; smallpox, 2-3 
Ware smallpox epidemic, 1722, 3 
Warlomont : admixture of gly- 
cerine with lymph patented in 
England by, 158 ; animal vac- 
cination introduced to Brussels 
by, 145 
Warrington smallpox epidemic, 

1773. 3 

Watson, Sir Thomas ; inocula- 
tion, 26-27 

Wegefarth ; protozoa, 124-125 

Wilson Hake ; analyses of gly- 
cerine, 172-173 

Woitow ; micro - organisms in 
vaccine lymph, 94-95 

Woodville ; arm-to-arm vaccina- 
tion, 68-70 

Yeasts, certain, action of glycerine 
on ; Blaxall and Copeman, 164- 

Zurn ; granules in vaccine lymph, 


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jWNDINQ ■SEC/. AUG 3 11965 

RC Copeman, Sydney Monckton 
183 Vaccination 


& Medical